Isolation of Pure Mitochondria from Rat Kidneys and WesternBlot of Mitochondrial Respiratory Chain Complexes
Cardiac, neuronal and renal tubular epithelial cells are the most metabolically active cells in the body. Their fate depends largely on their mitochondria as the primary energy generating system which participates in the control of apoptosis, cell cycle and metabolism. Thus, mitochondrial dysfunction is a hallmark of many chronic diseases including diabetic nephropathy. A drop in mitochondrial bioenergetics efficiency is often associated with altered expression of respiratory chain complexes. Moreover, recent studies demonstrate that cellular proteins can shuttle to mitochondria and modify their function directly.
Here we illustrate two mitochondria isolation protocols; one is recommended if the purity of the mitochondrial fraction is a priority such as if the mitochondrial localization of a protein has to be validated, the other if a high yield of intact functional mitochondria is required for functional studies and quantitative Western blotting. Next, we provide a detailed protocol for Western blotting of isolated mitochondria and renal cortex either to prove the purity of isolated fractions or to quantify complexes of the mitochondrial respiratory chain. We used this approach to identify classically cell membrane bound angiotensin II receptors in mitochondria and to study the effect of these receptors on mitochondrial function in early stages of diabetic nephropathy.
Identification of early diagnostic antigens in soluble proteins of Trichinella spiralis adult worms by Westernblot
Previous studies showed that crude antigens from Trichinella spiralis adult worms (AW) can be recognized by mouse infection sera at 8 days post infection. The aim of this study was to identify the early diagnostic antigenic bands in soluble proteins from T. spiralis AW by Western blot using early infection sera. The affecting factors of adult recovery were firstly observed in this study, and the results showed that the maximum number of adults was collected from small intestine when the female BALB/c mice were orally infected with 4000 ML and sacrificed at 3 days post infection.
The results of Western blot analysis showed that seven protein bands (31, 35.1, 39, 40.6, 41.9, 47 and 50.6 kDa) could be recognized by early infection sera as early as at 8-10 days post infection, and were strongly reacted with mouse infection sera at 11-12 days post infection. Our results suggested that the seven protein bands of T. spiralis AW soluble proteins might be the early expressed antigens during the intestinal stage of Trichinella infection and therefore have potential value for the early diagnosis of trichinellosis.
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.
Description: Western blot detection of IP proteins can exhibit high background due to the use of the same primary antibody in the IP and the Western blot. The Pure-IP Western Blot Detection Kit eliminates this background problem by using a proprietary HRP Conjugate for detection of the primary antibody.
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.
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.
Description: IFN beta is a mammalian Type I inferferon, functionig as a regulator of cellular activity by interacting with cell-surface receptors and activating various signaling pathways. IFN beta produces antiviral, antibacterial, and anticancer properties. Equine IFN beta Recombinant Protein is purified IFN beta produced in yeast.
Chicken Serum Albumin protein control for Western blot
Description: Chemokine ligand 3 (CCL3), also known as Macrophage Inflammatory Protein-1 alpha (MIP-1 alpha), is a small cytokine belonging to the CC chemokine family. CCL3 (MIP-1 alpha) is a chemoattractant for several different leukocytes, with varying degrees of potency. Equine CCL3 Recombinant Protein is purified chemokine ligand 3 (CCL3, MIP-1 alpha) produced in yeast.
Description: Erythropoietin (EPO) is an essential hormone for red blood cell production. Erythropoietin has a range of actions including vasoconstriction-dependent hypertension, stimulating angiogenesis, and inducing proliferation of smooth muscle fibers. Equine Erythropoietin Recombinant Protein is purified Erythropoietin (EPO) produced in yeast.
Description: CXCL9 (MIG) is a T-cell chemoattractant. Induced by IFN-gamma (IFN-γ), the ELR-negative chemokine CXCL9 (MIG) elicits its effects by binding to the cell surface chemokine receptor CXCR3. Equine CXCL9 Recombinant Protein is purified CXCL9 (MIG) produced in yeast.
Description: Chemokine ligand 11 (CCL11) belongs to the CC chemokine family and is commonly known as Eotaxin-1. CCL11 (Eotaxin-1) selectively recruits eosinophils by inducing their chemotaxis, and therefore, is implicated in allergic responses. Equine CCL11 Recombinant Protein is purified chemokine ligand 11 (CCL11, Eotaxin-1) produced in yeast.
Recombinant Norovirus 1 (MNV-1) Capsid Protein 1 (VP1) control for Western blot
Simple Western: Bringing the WesternBlot into the Twenty-First Century
The Western blot is widely used in the study of protein biochemistry, but it is notoriously labor-intensive, and it is limited in its reproducibility and quantification, among many other challenges. By contrast, capillary-based protein separation and immunodetection, known as Simple Western™, overcomes many of the challenges associated with the traditional Western blot, and it is quickly gaining traction as a replacement for traditional Western blot analysis.
The advantages that capillary-based immunoassay offers include ease of use, automation, reproducibility, quantification, and even built-in total protein normalization. In this chapter, we describe protocols for the two basic types of capillary-based immunodetection assays, one by molecular weight separation and the other by charge separation. In both methods, protein samples are separated in the capillary followed seamlessly by immunodetection with chemiluminescent or fluorescent antibodies for highly sensitive and specific detection of target proteins.
Biological Validation of a Novel Process and Product for Quantitating WesternBlots
Protein normalization of western blots has relied upon housekeeping proteins which exhibit signal saturation and varied cellular expression level variations. These issues can produce spurious results leading to erroneous conclusions. A superior method to protein normalization using housekeeping proteins is Total Protein Normalization, a method now recognized as the gold standard for quantitative westerns.
Total Protein Normalization requires that all proteins on a membrane be stained or labeled uniformly, imaged, and then analyzed for total protein. It is important that such a normalization process not interfere with typical immunodetection methods, fits within existing western workflows, and exhibits a linear relationship of signal intensity to protein load under all experimental conditions. Here we report that we developed a new reagent enabling Total Protein Normalization, and we demonstrate its superior protein normalization capabilities through analysis of target proteins in different cell backgrounds.
These data illustrate how housekeeping proteins exhibit signal saturation, yield erroneous normalization data, and display sample-to-sample variations averaging 48.2% overall. Signal intensities obtained using our new method show a linear relationship to protein sample load, thus providing accurate protein normalization with an overall average variation of 7.7%.
Identification of Protein Carbonyls (PCOs) in Canine Serum by WesternBlot Technique and Preliminary Evaluation of PCO Concentration in Dogs With Systemic Inflammation
In people, serum Protein Carbonyls (PCOs) increase during oxidative stress (OS) due to oxidative damage to proteins. OS is often associated with inflammation and especially with sepsis, a condition hard to diagnose in veterinary medicine because reliable markers are lacking. The aim of this study was to assess whether PCOs in canine serum may be detected by antibody-based methods such as Western Blotting (WB), and to preliminarily investigate the possible utility of this marker in dogs with inflammation.
A serum sample oxidized in vitro was used to set up the method; the coefficient of variation obtained by repeated analysis varied from 24 to 36%. In order to assess whether the technique may cover the range of PCOs concentration detectable in routine practice, PCOs were measured in 4 healthy dogs and in 15 with inflammatory diseases, in some cases potentially associated with sepsis, as suggested by the results of other inflammatory markers such as C-Reactive Protein (CRP) and the anti-oxidant enzyme Paraoxonase 1 (PON-1): the concentration of PCOs was low in dogs with normal PON-1 activity,
moderately increased in the majority of dogs with low-normal PON-1 activity, and severely increased in dogs with very low PON-1 activity. In conclusion this study demonstrates that PCOs, may be detected in canine serum, using antibody-based techniques such as WB. The preliminary results in dogs with and without systemic inflammation encourage further studies on the possible role of PCOs as inflammatory markers.
)
ChonBlock Western Blot Buffer 1 (TBS) 
TMB Substrate, for Western Blot 
Interferon alpha Western Blot Antibody 
AgileBlotter? Automated Western Blot Processor 
Hypersensitive Western Blot Chemiluminescent Substrate )
PDCD4 Western Blot kit (AWBK09045) )
ETS1 Western Blot kit (AWBK100604) )
ERG Western Blot kit (AWBK100625) )
GABRP Western Blot kit (AWBK13034) )
HTR1A Western Blot kit (AWBK13041) )
RAB14 Western Blot kit (AWBK13107) )
TNRC4 Western Blot kit (AWBK31525) )
ESRRG Western Blot kit (AWBK31655) )
GLI2 Western Blot kit (AWBK31885) )
PAX4 Western Blot kit (AWBK32064) )
AHR Western Blot kit (AWBK32243) )
GLI1 Western Blot kit (AWBK32368) )
PAX7 Western Blot kit (AWBK32393) )
LHX6 Western Blot kit (AWBK32553) )
HIPK2 Western Blot kit (AWBK32586) )
PAX7 Western Blot kit (AWBK32742) )
ZFP67 Western Blot kit (AWBK32749) )
OLIG2 Western Blot kit (AWBK32753) )
ENG Western Blot kit (AWBK33068) )
SOX5 Western Blot kit (AWBK33323) )
SOX10 Western Blot kit (AWBK33326) )
RPS16 Western Blot kit (AWBK33535) )
ATP7A Western Blot kit (AWBK33797) )
A1BG Western Blot kit (AWBK33810) )
NPM1 Western Blot kit (AWBK34094) )
LIG4 Western Blot kit (AWBK34122) )
RNF2 Western Blot kit (AWBK34290) )
ZFP36 Western Blot kit (AWBK34385) )
ARIH2 Western Blot kit (AWBK34418) )
KEAP1 Western Blot kit (AWBK34728) )
CLIC1 Western Blot kit (AWBK35045) )
KCNN2 Western Blot kit (AWBK35094) )
CLCN3 Western Blot kit (AWBK35504) )
P2RX7 Western Blot kit (AWBK35518) )
TRPM3 Western Blot kit (AWBK35609) )
PITX2 Western Blot kit (AWBK35634) )
ILF2 Western Blot kit (AWBK35731) )
ONECUT2 Western Blot kit (AWBK35750) )
ZFP42 Western Blot kit (AWBK36244) )
ANXA1 Western Blot kit (AWBK36568) )
SPP1 Western Blot kit (AWBK36677) )
RUNX2 Western Blot kit (AWBK36678) )
ZBP1 Western Blot kit (AWBK36711) )
Bapx1 Western Blot kit (AWBK36826) )
G3BP Western Blot kit (AWBK37713) )
TWIST1 Western Blot kit (AWBK37997) )
ID4 Western Blot kit (AWBK38058) )
SATB1 Western Blot kit (AWBK38208) )
EED Western Blot kit (AWBK38384) )
TARDBP Western Blot kit (AWBK38941) )
TARDBP Western Blot kit (AWBK38942) )
BRD7 Western Blot kit (AWBK39018) )
JMJD3 Western Blot kit (AWBK40102) )
SNRP70 Western Blot kit (AWBK40439) )
SSB Western Blot kit (AWBK40462) )
STAU1 Western Blot kit (AWBK40537) )
PCBP1 Western Blot kit (AWBK40630) )
HNRPLL Western Blot kit (AWBK41102) )
MSI2 Western Blot kit (AWBK41112) )
HNRPA3 Western Blot kit (AWBK41195) )
WNT2B Western Blot kit (AWBK41254) )
GNB1L Western Blot kit (AWBK41328) )
SLC12A1 Western Blot kit (AWBK41388) )
TPM1 Western Blot kit (AWBK41392) )
CEACAM6 Western Blot kit (AWBK41504) )
SLC22A1 Western Blot kit (AWBK41516) )
STK11 Western Blot kit (AWBK41743) )
OTC Western Blot kit (AWBK41766) )
TSHR Western Blot kit (AWBK41870) )
RHOBTB1 Western Blot kit (AWBK41883) )
KRT13 Western Blot kit (AWBK42031) )
IDH3A Western Blot kit (AWBK42237) )
SLC1A5 Western Blot kit (AWBK42247) )
PBEF1 Western Blot kit (AWBK42255) )
FST Western Blot kit (AWBK42287) )
SLC22A7 Western Blot kit (AWBK42306) )
ASPN Western Blot kit (AWBK42487) )
RSAD2 Western Blot kit (AWBK42696) )
PANX2 Western Blot kit (AWBK42778) )
PANX1 Western Blot kit (AWBK42783) )
TGFBI Western Blot kit (AWBK44268) )
PTPN2 Western Blot kit (AWBK45362) )
RORA Western Blot kit (AWBK45608) )
Chemiluminescent Western Blot Kit (OKRA00001) )
Chemiluminescent Western Blot Kit (OKRA00002) )
Chemiluminescent Western Blot Kit (OKRA00003) )
Chemiluminescent Western Blot Kit (OKRA00004) )
Colorimetric Western Blot Kit (OKRA00014) )
Colorimetric Western Blot Kit (OKRA00015) )
Colorimetric Western Blot Kit (OKRA00016) )
Colorimetric Western Blot Kit (OKRA00017) 
Western equine encephalitis PCR kit 
T-Pro Western Blot Stripping Reagent 
Western Blot Primary Antibody Diluent Buffer 
Attoglow Western Blot Analysis Kit:Blocking Agent 
Pure-IP Western Blot Detection Kit )
cyclin G2 Western Blot kit (AWBK03032) 
Bovine DNAse 1 Protein Western blot +ve control 
Mouse Leptin protein Western blot +ve control # 1 
Western equine encephalitis RT PCR kit 
Total Protein Western Blots - Human Adult Normal Tissue, Blot IV, 16 lanes )
Pre-made Minicircle CMV-GFP-SV40PolyA (positive control) )
Pre-made Minicircle CMV-rRFP-SV40PolyA (positive control)  Western blot +ve control)
Purified Ram Cyclooxygenase (Cox-1) Western blot +ve control 
West-Ez Blocking Buffer, 1 % Casein for Western Blot 
Poly-ADP-ribosylated proteins for Western blot 
Mouse Ceruloplasmin protein control for western blot 
Rat Ceruloplasmin protein control for western blot 
Attoglow Western Blot Analysis Kit:Binding Buffer 20x 
Recombinant Human MutY Protein for Western blot 
Nitrated Mol Wt Std for Western blot 
SuperBrite? ECL Western Blot Substrate/Detection Kit 
Western equine encephalitis One-Step PCR kit 
Custom Neg Control siRNA 
Custom Pos Control siRNA 
Equine IFN beta 1 Recombinant Protein 
Chicken Serum Albumin protein control for Western blot  Protein for Western Blot)
Beta-Galactosidase (E. Coli) Protein for Western Blot 
Recombinant Beta Lactamase protein control for Western blot 
Human C1 Esterase Inhibitor western blot +ve control 
Purified Human C1q protein Western blot +ve control 
Human Spleen Ferritin protein Western blot +ve control 
Human Liver Ferritin protein Western blot +ve control 
Horse Spleen Ferritin protein Western blot +ve control )
Western blot recycling stripping buffer, 50 ml (10X) )
ChonBlock Western Blot Buffer 2 for Goat (PBS) )
ChonBlock Western Blot Buffer 2 for Goat (TBS) )
ChonBlock Western Blot Buffer 2 for Rabbit (PBS) )
ChonBlock Western Blot Buffer 2 for Rabbit (TBS) 
Purified Cholera Toxin protein control for Western Blot 
Human Pancreatic Amylase protein control for Western blot 
Human Leptin protein Western blot +ve control # 2 )
HRP Western Blot Anti-IgG Antibody Kit (OKRA00006) 
Custom Peptide Synthesis order form  Protein control for western blot)
Recombinant 1 Envelope glycoprotein 2 (1 E2) Protein control for western blot 
Mouse NOS-II/i-NOS Protein Western Blot +ve Control # 1 
Equine CCL3 Recombinant Protei 
Equine EPO Recombinant Protein 
Equine CXCL9 Recombinant Protein 
Equine CCL11 Recombinant Protein  Capsid Protein 1 (VP1) control for Western blot)
Recombinant Norovirus 1 (MNV-1) Capsid Protein 1 (VP1) control for Western blot 
Purified Mouse serum albumin protein control for Western blot 
Purified Human serum albumin protein control for Western blot 
Purified rat serum albumin protein control for Western blot 
Purified Sheep serum albumin protein control for Western blot 
Human recombinant, purified BMP2 protein Western blot +ve control 
Human recombinant, purified BMP3 protein Western blot +ve control 
Human recombinant, purified BMP4 protein Western blot +ve control 
Human recombinant, purified BMP5 protein Western blot +ve control 
Human recombinant, purified BMP6 protein Western blot +ve control 
Recombinant human BAFF-R protein control for Western blot 
Human Adipsin/Factor D protein Western blot +ve control  protein control for western blot)
Human hemoglobin (non-glycated) protein control for western blot  solution for western blot)
Single solution TMB substrate (membrane) solution for western blot 
Western blot Kit for Rabbit Primary Antibodies, Chemilum. Substrate 
Western blot Kit for Goat Primary Antibodies, Chemilum. Substrate 
Western blot Kit for Mouse Primary Antibodies, Chemilum. Substrate 
Western blot Kit for Rat Primary Antibodies, Chemilum. Substrate 
Western blot Kit for Chicken Primary Antibodies, Chemilum. Substrate 
Western blot Kit for Monkey Primary Antibodies, Chemilum. Substrate 
Western blot Kit for Human Primary Antibodies, Chemilum. Substrate 
Western blot Kit for Sheep Primary Antibodies; Chemilum. Substrate 
Western blot Kit for Pig Primary Antibodies; Chemilum. Substrate 
NuGlo ECL Substrate Kit for Western blot, 400 mls 
WesternOne? Marker Pen, HRP for Western blot chemiluminescent detection 
Human Liver Cathepsin B protein control for Western blot 
Human Liver Cathepsin D protein control for Western blot 
Human Liver Cathepsin G protein control for Western blot 
Human Liver Cathepsin H protein control for Western blot 
