Identification of novel fibronectin-binding proteins by 2D-far Western blot

Identification of novel fibronectin-binding proteins by 2D-far Western blot

BirA⋆-protein A fusion protein primarily based BioEnhancer amplifies Western blot immunosignal

Western blot (protein immunoblot) is a extensively used analytical approach in molecular biology. Using the precise recognizing major antibody, proteins immobilized on numerous matrix are investigated by subsequent visualization steps, for instance, by the HRP conjugated secondary antibody incubation. Strategies to enhance the sensitivity in protein identification or quantification are appreciated by biochemists. Herein, we report a brand new technique to amplify Western blot indicators by establishing a probe with proximal labeling and IgG focusing on skills. The R118G mutation attenuated the biotin-AMP binding affinity of the bacterial biotin ligase BirA⋆, providing a proximity-dependent labeling means, which could possibly be used as a sign amplifier. We constructed a BirA⋆-protein A fusion protein (BioEnhancer) which particularly binds to IgG and provides biotin tags to its proximal amine teams, enhancing the immunosignal of goal proteins. In our experiments, the BioEnhancer system amplified the immunosignal by 10-fold in comparison with the usual Western blot. Moreover, our technique might couple with different sign enhancement strategies to additional improve the Western blot sensitivity. This text is protected by copyright. All rights reserved.

 

A quantitative western blot approach utilizing TMB: Comparability with the standard approach

Quite a few molecular organic experiments carried out all through the world require the detection or quantification of a protein of curiosity. Western blotting is among the hottest methods used for this goal and affords quantitative info with assistance from specialised software program. Nonetheless, its dependence on the image that’s captured, and the background and the absence of a typical protocol stop the approach from being fully quantitative.

To beat these obstacles, we current a easy and dependable assay that’s much like the common approach, except the final stage of band visualization and quantification. We suggest that small items of the blot that embody the protein of curiosity could be minimize and dipped in a small quantity of three,3′,5,5′-Tetramethylbenzidine (TMB) resolution, giving a colorimetric sign with linear dependence on the amount of the protein.

The response is stopped with H2 SO4 , and the sign is measured in a plate reader. This modification exhibits excessive linearity with out extra prices and could be utilized for each purified proteins and proteins present in a lysate. The outcomes obtained with our proposed approach had been in contrast with these obtained by the standard technique and proved to be extra dependable.

nwreprosci
nwreprosci

WESTSAVE Up (Western Blotting Substrate)

LF-QC0101 1 kit
EUR 181.2
Description: Abfrontier WESTSAVE UpTM (Western Blotting Substrate) is enhanced luminol-based chemiluminescent substrate for the non-radioactive detection of Horseradish Peroxidase(HRP) labeled antibodies.

ECL PRIME WESTERN BLOTTING SYSTEM

GERPN2232 EACH
EUR 254.22

ECL Prime Western Blotting Det

RPN2232 EACH
EUR 258.78

Amersham ECL DualVue Western Blotting Markers

RPN810 EACH
EUR 223.44

WESTSAVE Up (Western Blotting Substrate, double pack)

LF-QC0102 1 kit(Double pack)
EUR 266.4
Description: Abfrontier WESTSAVE UpTM (Western Blotting Substrate) is enhanced luminol-based chemiluminescent substrate for the non-radioactive detection of Horseradish Peroxidase(HRP) labeled antibodies.

GloBrite chemiluminescence reagent kit for western blotting 200 mL

GLB1 200 mL
EUR 174

GloBrite chemiluminescence reagent kit for western blotting 500 mL

GLB2 500 mL
EUR 280.8

Western Blotting Mouse IgG DAB Chromogenic Reagent Kit (Yellow)

SA2020 1 kit(1200 cm2)
EUR 169.2

Western Blotting Goat IgG DAB Chromogenic Reagent Kit (Yellow)

SA2021 1 kit(1200 cm2)
EUR 169.2

Western Blotting Rabbit IgG DAB Chromogenic Reagent Kit (Yellow)

SA2022 1 kit(1200 cm2)
EUR 169.2

Western Blotting Rat IgG DAB Chromogenic Reagent Kit (Yellow)

SA2023 1 kit(1200 cm2)
EUR 169.2

Western Blotting Mouse IgG DAB Chromogenic Reagent Kit (Blue)

SA2024 1 kit(1200 cm2)
EUR 169.2

Western Blotting Rabbit IgG DAB Chromogenic Reagent Kit (Blue)

SA2025 1 kit(1200 cm2)
EUR 169.2

Western Blotting Filter Paper, 12.5 cm—12.5 cm

AR0172 100 sheets
EUR 84

Western Blotting Filter Paper, 9 cm— 7.5 cm

AR0173 100 sheets
EUR 78

Purified Lassa fever Nucleoprotein control for Western Blotting

LFNP15-C-10 100 ul
EUR 343.2

Purified Nipah virus Glycoprotein control for Western Blotting

NIV11-C 100 ul
EUR 343.2

Purified Nipah virus Nucleoprotein control for Western Blotting

NIV21-C 100 ul
EUR 270

Mayaro virus (MAYV) Capsid Protein control for Western Blotting

MAYV31-C 100 ul
EUR 343.2

Mayaro virus (MAYV) nsP1 protein control for Western Blotting

MAYV41-C 100 ul
EUR 343.2

Mayaro virus (MAYV) 6K Protein control for Western Blotting

MAYV51-C 100 ug
EUR 343.2

Mayaro virus (MAYV) E3 protein control for Western Blotting

MAYV61-C 100 ul
EUR 343.2

Amersham ECL Western Blotting Detection Reagents for 4000cm2 membrane

RPN2106 EACH
EUR 314.64

Amersham ECL Western Blotting Detection Reagents for 1000cm2 membrane

RPN2109 EACH
EUR 158.46

Amersham ECL Western Blotting Detection Reagents for 6000cm2 membrane

RPN2134 1KIT
EUR 482.22

Amersham ECL Western Blotting Detection Reagents for 2000cm2 membrane

RPN2209 EACH
EUR 245.1

Human Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

DLR-KEAP1-Hu-48T 48T
EUR 664.8
Description: A sandwich quantitative ELISA assay kit for detection of Human Kelch Like ECH Associated Protein 1 (KEAP1) in samples from serum, plasma, tissue homogenates or other biological fluids.

Human Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

DLR-KEAP1-Hu-96T 96T
EUR 870
Description: A sandwich quantitative ELISA assay kit for detection of Human Kelch Like ECH Associated Protein 1 (KEAP1) in samples from serum, plasma, tissue homogenates or other biological fluids.

Mouse Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

DLR-KEAP1-Mu-48T 48T
EUR 679.2
Description: A sandwich quantitative ELISA assay kit for detection of Mouse Kelch Like ECH Associated Protein 1 (KEAP1) in samples from serum, plasma, tissue homogenates or other biological fluids.

Mouse Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

DLR-KEAP1-Mu-96T 96T
EUR 889.2
Description: A sandwich quantitative ELISA assay kit for detection of Mouse Kelch Like ECH Associated Protein 1 (KEAP1) in samples from serum, plasma, tissue homogenates or other biological fluids.

Rat Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

DLR-KEAP1-Ra-48T 48T
EUR 710.4
Description: A sandwich quantitative ELISA assay kit for detection of Rat Kelch Like ECH Associated Protein 1 (KEAP1) in samples from serum, plasma, tissue homogenates or other biological fluids.

Rat Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

DLR-KEAP1-Ra-96T 96T
EUR 932.4
Description: A sandwich quantitative ELISA assay kit for detection of Rat Kelch Like ECH Associated Protein 1 (KEAP1) in samples from serum, plasma, tissue homogenates or other biological fluids.

Rat Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

RD-KEAP1-Ra-48Tests 48 Tests
EUR 727.2

Rat Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

RD-KEAP1-Ra-96Tests 96 Tests
EUR 1012.8

Human Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

RDR-KEAP1-Hu-48Tests 48 Tests
EUR 706.8

Human Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

RDR-KEAP1-Hu-96Tests 96 Tests
EUR 984

Mouse Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

RDR-KEAP1-Mu-48Tests 48 Tests
EUR 723.6

Mouse Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

RDR-KEAP1-Mu-96Tests 96 Tests
EUR 1008

Rat Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

RDR-KEAP1-Ra-48Tests 48 Tests
EUR 759.6

Rat Kelch Like ECH Associated Protein 1 (KEAP1) ELISA Kit

RDR-KEAP1-Ra-96Tests 96 Tests
EUR 1060.8

Amersham ECL Western Blotting Systemor detection for 1000 cm2 membrane

RPN2108 EACH
EUR 274.74

Amersham ECL Plex Western Blotting Combination Pack Cy3 Cy5 HybondLFP

RPN999 EACH
EUR 235.98

Keap1/ Rat Keap1 ELISA Kit

ELI-38983r 96 Tests
EUR 1063.2

Amersham ECL Plex Western Blotting Combination Pack Cy3 Cy5 Hybond ECL

RPN998 1KIT
EUR 256.5

Blotting roller

2322479 4unit
EUR 320.4

Ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco/Rbcl) Loading Control for Western Blotting

RBCL15-C 100 ul
EUR 343.2

Blotting Paper Sheets

F3030-6188 PK100
EUR 57

Western Blotting Boxes, 6-Compartment, 1 3/8 Height ( 32x100 mm Each component)

LC7012-004 4/Pk
EUR 116.4

Western Blot Kit

20-abx098123
  • EUR 276.00
  • EUR 343.20
  • 100 ml
  • 200 ml

Western Blotting Boxes,15.2 x 10.2 x 5.1 cm (6 x 4 x 2in)

LC7021-005 5/Pk
EUR 102

KEAP1 ELISA KIT|Human

EF007411 96 Tests
EUR 826.8

KEAP1 Antibody

1-CSB-PA247380
  • EUR 380.40
  • EUR 292.80
  • 100ul
  • 50ul
Description: A polyclonal antibody against KEAP1. Recognizes KEAP1 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, IHC;ELISA:1:2000-1:5000, IHC:1:25-1:100

KEAP1 Antibody

1-CSB-PA005148
  • EUR 266.40
  • EUR 234.00
  • 100ug
  • 50ug
Description: A polyclonal antibody against KEAP1. Recognizes KEAP1 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: WB, ELISA;WB:1/500-1/2000.ELISA:1/20000

Keap1 Antibody

DF6637 200ul
EUR 420

Keap1 Antibody

AF5266 200ul
EUR 420

KEAP1 Antibody

1-CSB-PA011976
  • EUR 380.40
  • EUR 292.80
  • 100ul
  • 50ul
Description: A polyclonal antibody against KEAP1. Recognizes KEAP1 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, IHC;ELISA:1:2000-1:5000, IHC:1:25-1:100

KEAP1 Antibody

1-CSB-PA012147GA01HU
  • EUR 716.40
  • EUR 399.60
  • 150ul
  • 50ul
Description: A polyclonal antibody against KEAP1. Recognizes KEAP1 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC

KEAP1 Antibody

1-CSB-PA012147LA01HU
  • EUR 380.40
  • EUR 402.00
  • 100ug
  • 50ug
Description: A polyclonal antibody against KEAP1. Recognizes KEAP1 from Human, Mouse. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC; Recommended dilution: WB:1:200-1:1000, IHC:1:20-1:200

Keap1 Antibody

1-CSB-PA012147LA01MO
  • EUR 380.40
  • EUR 402.00
  • 100ug
  • 50ug
Description: A polyclonal antibody against Keap1. Recognizes Keap1 from Human, Mouse. This antibody is Unconjugated. Tested in the following application: ELISA, WB; Recommended dilution: WB:1:500-1:2000

KEAP1 antibody

70R-8050 50 ug
EUR 560.4
Description: Affinity purified rabbit polyclonal KEAP1 antibody

KEAP1 antibody

70R-8295 50 ug
EUR 560.4
Description: Affinity purified rabbit polyclonal KEAP1 antibody

KEAP1 Antibody

32450-100ul 100ul
EUR 302.4

KEAP1 Antibody

42790-100ul 100ul
EUR 302.4

KEAP1 Antibody

7045-002mg 0.02 mg
EUR 206.18
Description: KEAP1 Antibody: KEAP1 (kelch-like ECH-associated protein 1) is a stress sensing adaptor for the Cullin3 (Cul3)-dependent E3 ubiquitin ligase complex that negatively regulates NRF2 (NF-E2-related factor 2) and plays a role in the oxidative stress response. It targets NFE2L2/NRF2 for ubiquitination and degradation by the proteasome. KEAP1 contains an amino terminal BTB/POZ domain and a carboxyl terminal KELCH domain which are required for interaction with NRF2, and in binding Cul3-E3 ubiquitin ligase. Altered expression of NRF2 is associated with chronic obstructive pulmonary disease (COPD). KEAP1 also targets the down regulation of NF-κB activity by targeting IKKβ degradation. Mutation of the KEAP1 gene is found in lung cancer.

KEAP1 Antibody

7045-01mg 0.1 mg
EUR 523.7
Description: KEAP1 Antibody: KEAP1 (kelch-like ECH-associated protein 1) is a stress sensing adaptor for the Cullin3 (Cul3)-dependent E3 ubiquitin ligase complex that negatively regulates NRF2 (NF-E2-related factor 2) and plays a role in the oxidative stress response. It targets NFE2L2/NRF2 for ubiquitination and degradation by the proteasome. KEAP1 contains an amino terminal BTB/POZ domain and a carboxyl terminal KELCH domain which are required for interaction with NRF2, and in binding Cul3-E3 ubiquitin ligase. Altered expression of NRF2 is associated with chronic obstructive pulmonary disease (COPD). KEAP1 also targets the down regulation of NF-κB activity by targeting IKKβ degradation. Mutation of the KEAP1 gene is found in lung cancer.

KEAP1 Peptide

7045P 0.05 mg
EUR 197.7
Description: (CT) KEAP1 peptide

KEAP1 antibody

70R-18096 50 ul
EUR 522
Description: Rabbit polyclonal KEAP1 antibody

KEAP1 antibody

70R-14195 100 ug
EUR 418.8
Description: Affinity purified Rabbit polyclonal KEAP1 antibody

KEAP1 Peptide

42-391P 0.1 mg
EUR 405.6
Description: (IN) (near (NT) KEAP1 Peptide

KEAP1 antibody

20R-1096 100 ug
EUR 452.4
Description: Rabbit polyclonal KEAP1 antibody

KEAP1 antibody

10R-7009 100 ul
EUR 871.2
Description: Mouse monoclonal KEAP1 antibody

KEAP1 antibody

10R-4529 100 ul
EUR 829.2
Description: Mouse monoclonal KEAP1 antibody

KEAP1 antibody

10R-4530 100 ul
EUR 829.2
Description: Mouse monoclonal KEAP1 antibody

Keap1 Antibody

ABF5266 100 ug
EUR 525.6

KEAP1 Antibody

ABD6637 100 ug
EUR 525.6

KEAP1 siRNA

20-abx921425
  • EUR 661.20
  • EUR 878.40
  • 15 nmol
  • 30 nmol

KEAP1 siRNA

20-abx921426
  • EUR 661.20
  • EUR 878.40
  • 15 nmol
  • 30 nmol

KEAP1 Antibody

BF0010 200ul
EUR 540

KEAP1 Antibody

R32225 100 ug
EUR 419

KEAP1 Antibody

R34041-100UG 100 ug
EUR 399

KEAP1 Antibody

F40314-0.08ML 0.08 ml
EUR 165

KEAP1 Antibody

F40314-0.4ML 0.4 ml
EUR 379

KEAP1 Antibody

F40464-0.08ML 0.08 ml
EUR 165

KEAP1 Antibody

F40464-0.4ML 0.4 ml
EUR 379

Frit Kit

FRIT-KIT 1each
EUR 148.8
Description: Kit to create frits in capillaries. Includes formamide, Kasil-1, Kasil-1624 and a cleaving tool.

Western Blot Enhancing Kit

K3172120 1200 cm2
EUR 171.6
Description: Can be used for various proteomics studies in both normal and pathological cases. It is an excellent control and suitable for educational purposes. This product is prepared from whole tissue homogenates and has undergone SDS-PAGE quality control analysis. The protein is stored in a buffer with protease inhibitor cocktail fo prevent degradation.

Western Blot Enhancing Kit

K3172250 2500 cm2
EUR 225.6
Description: Can be used for various proteomics studies in both normal and pathological cases. It is an excellent control and suitable for educational purposes. This product is prepared from whole tissue homogenates and has undergone SDS-PAGE quality control analysis. The protein is stored in a buffer with protease inhibitor cocktail fo prevent degradation.

Column Packing Kit

PACK-KIT 1pack
EUR 1242
Description: Column packing kit for pressure cells. Includes: HPREG regulator, TBNG10 tubing, CAP-75 capillary, and STRB5X2 stir bar.

KEAP1 ELISA Kit (Rat) (OKCA01896)

OKCA01896 96 Wells
EUR 1015.2
Description: Description of target: Acts as a substrate adapter protein for the E3 ubiquitin ligase complex formed by CUL3 and RBX1 and targets NFE2L2/NRF2 for ubiquitination and degradation by the proteasome, thus resulting in the suppression of its transcriptional activity and the repression of antioxidant response element-mediated detoxifying enzyme gene expression. Retains NFE2L2/NRF2 and may also retain BPTF in the cytosol. Targets PGAM5 for ubiquitination and degradation by the proteasome.<p>This subsection of the <a href="//www.uniprot.org/help/function_section">‘Function’</a> section describes the metabolic pathway(s) associated with a protein.<p><a href='/help/pathway' target='_top'>More...</a></p>Pathwayi: protein ubiquitinationThis protein is involved in the pathway protein ubiquitination, which is part of Protein modification._x000D_View all proteins of this organism that are known to be involved in the pathway protein ubiquitination and in Protein modification. ;Species reactivity: Rat;Application: ;Assay info: Assay Methodology: Quantitative Sanadwich ELISA;Sensitivity: 11.75 pg/mL

KEAP1 ELISA Kit (Human) (OKCD00608)

OKCD00608 96 Wells
EUR 1090.8
Description: Description of target: Acts as a substrate adapter protein for the E3 ubiquitin ligase complex formed by CUL3 and RBX1 and targets NFE2L2/NRF2 for ubiquitination and degradation by the proteasome, thus resulting in the suppression of its transcriptional activity and the repression of antioxidant response element-mediated detoxifying enzyme gene expression. Retains NFE2L2/NRF2 and may also retain BPTF in the cytosol. Targets PGAM5 for ubiquitination and degradation by the proteasome.5 Publications <p>Manually curated information for which there is published experimental evidence.</p> <p><a href="/manual/evidences#ECO:0000269">More…</a></p> Manual assertion based on experiment iniRef.7"Distinct cysteine residues in Keap1 are required for Keap1-dependent ubiquitination of Nrf2 and for stabilization of Nrf2 by chemopreventive agents and oxidative stress."_x005F_x005F_x000D_Zhang D.D., Hannink M._x005F_x005F_x000D_Mol. Cell. Biol. 23:8137-8151(2003) [PubMed] [Europe PMC] [Abstract]Cited for: FUNCTION, MUTAGENESIS OF CYS-151; CYS-273 AND CYS-288, SUBCELLULAR LOCATION.Ref.8"Fetal Alz-50 clone 1 interacts with the human orthologue of the Kelch-like Ech-associated protein."_x005F_x005F_x000D_Strachan G.D., Morgan K.L., Otis L.L., Caltagarone J., Gittis A., Bowser R., Jordan-Sciutto K.L._x005F_x005F_x000D_Biochemistry 43:12113-12122(2004) [PubMed] [Europe PMC] [Abstract]Cited for: INTERACTION WITH NF2L2/NRF2 AND BPTF, FUNCTION, TISSUE SPECIFICITY, SUBCELLULAR LOCATION.Ref.9"Keap1 is a redox-regulated substrate adaptor protein for a Cul3-dependent ubiquitin ligase complex."_x005F_x005F_x000D_Zhang D.D., Lo S.-C., Cross J.V., Templeton D.J., Hannink M._x005F_x005F_x000D_Mol. Cell. Biol. 24:10941-10953(2004) [PubMed] [Europe PMC] [Abstract]Cited for: FUNCTION, INTERACTION WITH CUL3 AND RBX1, MUTAGENESIS OF 125-ILE--GLY-127 AND 162-TYR--ILE-164, UBIQUITINATION.Ref.10"Ubiquitination of Keap1, a BTB-Kelch substrate adaptor protein for Cul3, targets Keap1 for degradation by a proteasome-independent pathway."_x005F_x005F_x000D_Zhang D.D., Lo S.C., Sun Z., Habib G.M., Lieberman M.W., Hannink M._x005F_x005F_x000D_J. Biol. Chem. 280:30091-30099(2005) [PubMed] [Europe PMC] [Abstract]Cited for: FUNCTION, IDENTIFICATION IN A COMPLEX WITH CUL3 AND RBX1, UBIQUITINATION, ENZYME REGULATION, MUTAGENESIS OF CYS-151.Ref.13"PGAM5, a Bcl-XL-interacting protein, is a novel substrate for the redox-regulated Keap1-dependent ubiquitin ligase complex."_x005F_x005F_x000D_Lo S.-C., Hannink M._x005F_x005F_x000D_J. Biol. Chem. 281:37893-37903(2006) [PubMed] [Europe PMC] [Abstract]Cited for: INTERACTION WITH PGAM5, FUNCTION, ENZYME REGULATION, DOMAIN, MUTAGENESIS OF CYS-151; TYR-334; ARG-415; ARG-483 AND TYR-572. ;Species reactivity: Human;Application: ELISA;Assay info: Assay Methodology: Quantitative Sandwich Immunoassay;Sensitivity: < 0.056 ng/mL

KEAP1 ELISA Kit (Mouse) (OKCD00609)

OKCD00609 96 Wells
EUR 1123.2
Description: Description of target: Acts as a substrate adapter protein for the E3 ubiquitin ligase complex formed by CUL3 and RBX1 and targets NFE2L2/NRF2 for ubiquitination and degradation by the proteasome, thus resulting in the suppression of its transcriptional activity and the repression of antioxidant response element-mediated detoxifying enzyme gene expression. Retains NFE2L2/NRF2 and may also retain BPTF in the cytosol. Targets PGAM5 for ubiquitination and degradation by the proteasome (By similarity).By similarity2 Publications <p>Manually curated information for which there is published experimental evidence.</p> <p><a href="/manual/evidences#ECO:0000269">More…</a></p> Manual assertion based on experiment iniRef.1"Keap1 represses nuclear activation of antioxidant responsive elements by Nrf2 through binding to the amino-terminal Neh2 domain."_x005F_x005F_x000D_Itoh K., Wakabayashi N., Katoh Y., Ishii T., Igarashi K., Engel J.D., Yamamoto M._x005F_x005F_x000D_Genes Dev. 13:76-86(1999) [PubMed] [Europe PMC] [Abstract]Cited for: NUCLEOTIDE SEQUENCE [MRNA], FUNCTION.Ref.6"Keap1-dependent proteasomal degradation of transcription factor Nrf2 contributes to the negative regulation of antioxidant response element-driven gene expression."_x005F_x005F_x000D_McMahon M., Itoh K., Yamamoto M., Hayes J.D._x005F_x005F_x000D_J. Biol. Chem. 278:21592-21600(2003) [PubMed] [Europe PMC] [Abstract]Cited for: FUNCTION. <p>Describes the metabolic pathway(s) associated with a protein.</p><p><a href='../manual/pathway' target='_top'>More...</a></p>Pathwayi: protein ubiquitinationThis protein is involved in the pathway protein ubiquitination, which is part of Protein modification._x005F_x005F_x000D_View all proteins of this organism that are known to be involved in the pathway protein ubiquitination and in Protein modification. ;Species reactivity: Mouse;Application: ELISA;Assay info: Assay Methodology: Quantitative Sandwich Immunoassay;Sensitivity: < 0.055 ng/mL

KEAP1 ELISA Kit (Human) (OKEH08527)

OKEH08527 96 Wells
EUR 1075.2
Description: Description of target: This gene encodes a protein containing KELCH-1 like domains, as well as a BTB/POZ domain. Kelch-like ECH-associated protein 1 interacts with NF-E2-related factor 2 in a redox-sensitive manner and the dissociation of the proteins in the cytoplasm is followed by transportation of NF-E2-related factor 2 to the nucleus. This interaction results in the expression of the catalytic subunit of gamma-glutamylcysteine synthetase. Two alternatively spliced transcript variants encoding the same isoform have been found for this gene.;Species reactivity: Human;Application: ELISA;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 40.6pg/mL

Keap1 ELISA Kit (Rat) (OKEH08528)

OKEH08528 96 Wells
EUR 1075.2
Description: Description of target: ;Species reactivity: Rat;Application: ELISA;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 0.081ng/mL

Western Blotting Boxes, 15.2 x 10.2 x 3.2 cm (6 x 4 x 1 1/4in)

LC7020-005 5/Pk
EUR 100.8

PCR Mycoplasma Detection Kit

M034-Kit Kit
EUR 319.2

Blotting Paper, 10X15cm, 100 Sheets

PP3322 1PK, 100UNIT
EUR 126.82

Blotting Paper, 15X15cm, 100 Sheets

PP3323 1PK, 100UNIT
EUR 140.45

Blotting Paper, 20X20cm, 100 Sheets

PP3324 1PK, 100UNIT
EUR 199.68

Whatman GB005 Blotting Paper 58x58cm

MOL3119 PK25
EUR 394.44

3MM CHR Blotting Paper 12x14CM

F30306132 100PK
EUR 28.5

Gel Blotting Paper 20X20 Gb005

426981 PK25
EUR 96.9

Slide blotting paper TSC 89X140mm

TS10020 PK1000
EUR 67.26

Nonfat Dry Milk Blotting Grade

20-241 500g/Unit
EUR 272

KEAP1 cloning plasmid

CSB-CL012147HU-10ug 10ug
EUR 451.2
Description: A cloning plasmid for the KEAP1 gene.

Keap1 Polyclonal Antibody

ABP52949-003ml 0.03ml
EUR 189.6
Description: A polyclonal antibody for detection of Keap1 from Human, Mouse, Rat. This Keap1 antibody is for WB, IHC-P, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from the Internal region of human Keap1

Keap1 Polyclonal Antibody

ABP52949-01ml 0.1ml
EUR 346.8
Description: A polyclonal antibody for detection of Keap1 from Human, Mouse, Rat. This Keap1 antibody is for WB, IHC-P, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from the Internal region of human Keap1

Keap1 Polyclonal Antibody

ABP52949-02ml 0.2ml
EUR 496.8
Description: A polyclonal antibody for detection of Keap1 from Human, Mouse, Rat. This Keap1 antibody is for WB, IHC-P, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from the Internal region of human Keap1

KEAP1 Blocking Peptide

DF6637-BP 1mg
EUR 234

KEAP1 Conjugated Antibody

C32450 100ul
EUR 476.4

Keap1 Blocking Peptide

AF5266-BP 1mg
EUR 234

anti- KEAP1 antibody

FNab04516 100µg
EUR 606.3
Description: Antibody raised against KEAP1

anti- KEAP1 antibody

FNab04517 100µg
EUR 606.3
Description: Antibody raised against KEAP1

Keap1 Polyclonal Antibody

ES3948-100ul 100ul
EUR 334.8
Description: A Rabbit Polyclonal antibody against Keap1 from Human/Mouse/Rat. This antibody is tested and validated for WB, ELISA, IHC, WB, ELISA

Keap1 Polyclonal Antibody

ES3948-50ul 50ul
EUR 248.4
Description: A Rabbit Polyclonal antibody against Keap1 from Human/Mouse/Rat. This antibody is tested and validated for WB, ELISA, IHC, WB, ELISA

KEAP1 Rabbit pAb

A11258-100ul 100 ul
EUR 369.6

KEAP1 Rabbit pAb

A11258-200ul 200 ul
EUR 550.8

KEAP1 Rabbit pAb

A11258-20ul 20 ul
EUR 219.6

KEAP1 Rabbit pAb

A11258-50ul 50 ul
EUR 267.6

KEAP1 Rabbit pAb

A11484-100ul 100 ul
EUR 369.6

KEAP1 Rabbit pAb

A11484-200ul 200 ul
EUR 550.8

KEAP1 Rabbit pAb

A11484-20ul 20 ul
EUR 219.6

KEAP1 Rabbit pAb

A11484-50ul 50 ul
EUR 267.6

Keap1 Polyclonal Antibody

41626-100ul 100ul
EUR 302.4

Keap1 Polyclonal Antibody

41626-50ul 50ul
EUR 224.4

KEAP1 Blocking Peptide

33R-3861 100 ug
EUR 216
Description: A synthetic peptide for use as a blocking control in assays to test for specificity of KEAP1 antibody, catalog no. 70R-8295

KEAP1 Blocking Peptide

33R-8707 100 ug
EUR 216
Description: A synthetic peptide for use as a blocking control in assays to test for specificity of KEAP1 antibody, catalog no. 70R-8050

KEAP1 Blocking Peptide

33R-9382 100 ug
EUR 216
Description: A synthetic peptide for use as a blocking control in assays to test for specificity of KEAP1 antibody, catalog no. 20R-1096

Identification of novel fibronectin-binding proteins by 2D-far Western blot in atypical enteropathogenic Escherichia coli serotype O55:H7

Atypical enteropathogenic Escherichia coli (aEPEC) is a subgroup of EPEC, which is among the main pathogens liable for deadly diarrhoea in youngsters. In contrast with typical EPEC (tEPEC), aEPEC lack an EAF (EPEC adherence issue) plasmid (pEAF), which encodes a sequence of virulence-associated genes.

The extracellular matrix (ECM) element of human cells has been reported to be an vital component within the interplay between host and bacterial pathogens. On this analysis, a 2D-Far Western blot technique was carried out to identifiy the bacterial proteins that might bind to fibronectin, one of the vital frequent constituents of ECM. A complete of 17 protein spots had been recognized, together with Four outer membrane proteins (OMPs), specifically, OmpC, OmpD, OmpX and LamB. In vitro research had been used to find out whether or not these OMPs had been concerned within the adherence course of.

By way of oblique immunofluorescence assays, 4 OMPs could possibly be noticed on the surfaces of host cells. After incubating the cells with the recombinant proteins, the adhesion fee of the O55:H7 isolate was decreased. Moreover, the deletion of OmpX and LamB also can lower the adhesion fee of WT. Taken collectively, a high-throughput screening technique for host ECM-binding proteins primarily based on 2D Far-Western blot was established, and 4 outer membrane proteins recognized by this technique had been discovered to be concerned within the adherence course of.

Validation of superior reference genes for qRT-PCR and Western blot analyses in marine Emiliania huxleyi – virus mannequin system

Goals: To seek for a set of reference genes for dependable gene expression evaluation within the globally vital marine coccolithophore Emiliania huxleyi -virus mannequin system.

Strategies and outcomes: Fifteen housekeeping genes (CDKA, CYP15, EFG3, POLAI, RPL30, RPL13, SAMS, COX1, GPB1-2, HSP90, TUA, TUB, UBA1, CAM3 and GAPDH) had been evaluated for his or her stability as potential reference genes for qRT-PCR utilizing ΔCt, geNorm, NormFinder, Bestkeeper and RefFinder software program. CDKA, TUA and TUB genes had been examined as loading controls for Western blot in the identical pattern panel. Moreover, goal genes related to cell apoptosis, i.e., metacaspase genes, had been utilized to validate the collection of reference genes.

The evaluation outcomes demonstrated that putative housekeeping genes exhibited vital variations in each mRNA and protein content material throughout virus an infection. After a complete evaluation with all the algorithms, CDKA and GAPDH had been beneficial as probably the most secure reference genes for E huxleyi virus (EhV) an infection remedies. For Western blot, vital variation was seen for TUA and TUB, whereas CDKA was stably expressed, in keeping with the outcomes of qRT-PCR.

Conclusions: CDKA and GAPDH are the only option for gene and protein expression evaluation than the opposite candidate reference genes below EhV an infection situations.

Questioning protection values decided by 2D Western Blots – a essential examine on the characterization of anti-HCP ELISA reagents

 

Host cell proteins (HCPs) represent a serious class of process-related impurities, whose substantial clearance should be demonstrated by appropriate analytical strategies to warrant product high quality and cut back potential security dangers for sufferers. On this regard, enzyme linked immunosorbent assays (ELISAs), which primarily depend on the standard of the HCP reference normal (immunogen) and HCP-specific polyclonal antibodies, are thought of the gold normal for HCP monitoring.

For the qualification of the employed antibodies, 2D Western Blots (2D-WBs) are the popular approach to find out the protection, although quite a lot of sensible constraints are properly acknowledged. By utilizing a number of orthogonal approaches, corresponding to affinity-based mass spectrometry and oblique ELISA, the current examine revealed potential detection gaps (i.e., non-covered HCPs) of typical 2D-WBs, which could be primarily attributed to two completely different root causes: i) low quantities of proteins or antibodies being unable to beat the detection restrict and ii) western blot artifacts as a result of lack of conformational epitopes via protein denaturation hindering HCP-antibody recognition.

In distinction, the shortage of particular antibodies towards sure (significantly, low molecular weight) HCPs, as proposed in earlier research, appears to play solely a minor position. Collectively, these findings suggest that CHO-HCP ELISA antibodies are higher than qualification research by 2D-WBs point out. This text is protected by copyright. All rights reserved.

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