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Identification of early diagnostic antigens in soluble proteins of Trichinella spiralis
Bioassay Cultrex Enzyme Monoclonal Polyclonal Vector

Identification of early diagnostic antigens in soluble proteins of Trichinella spiralis

Isolation of Pure Mitochondria from Rat Kidneys and Western Blot 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 Western blot

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.

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Hypersensitive Western Blot Chemiluminescent Substrate
CR0001-5 10mL (sufficient reagents for 2000 cm2 of membrane)
EUR 116

AgileBlotter? Automated Western Blot Processor
ACT-ABH-40
EUR 16939

Interferon alpha Western Blot Antibody
20-abx137014
  • EUR 356.00
  • EUR 523.00
  • 0.5 mg
  • 1 mg

Chemiluminescent Western Blot Kit (OKRA00001)
OKRA00001 1 kit
EUR 570
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

Chemiluminescent Western Blot Kit (OKRA00002)
OKRA00002 1 kit
EUR 570
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

Chemiluminescent Western Blot Kit (OKRA00003)
OKRA00003 1 kit
EUR 570
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

Chemiluminescent Western Blot Kit (OKRA00004)
OKRA00004 1 kit
EUR 570
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

Colorimetric Western Blot Kit (OKRA00014)
OKRA00014 1 kit
EUR 609
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

Colorimetric Western Blot Kit (OKRA00015)
OKRA00015 1 each
EUR 609
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

Colorimetric Western Blot Kit (OKRA00016)
OKRA00016 1 kit
EUR 609
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

Colorimetric Western Blot Kit (OKRA00017)
OKRA00017 1 kit
EUR 609
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

PDCD4 Western Blot kit (AWBK09045)
AWBK09045 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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ETS1 Western Blot kit (AWBK100604)
AWBK100604 10 reactions
EUR 647
Description:
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  • Species reactivity:
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ERG Western Blot kit (AWBK100625)
AWBK100625 10 reactions
EUR 647
Description:
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  • Species reactivity:
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GABRP Western Blot kit (AWBK13034)
AWBK13034 10 reactions
EUR 647
Description:
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  • Species reactivity:
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HTR1A Western Blot kit (AWBK13041)
AWBK13041 10 reactions
EUR 647
Description:
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  • Species reactivity:
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RAB14 Western Blot kit (AWBK13107)
AWBK13107 10 reactions
EUR 647
Description:
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TNRC4 Western Blot kit (AWBK31525)
AWBK31525 10 reactions
EUR 647
Description:
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ESRRG Western Blot kit (AWBK31655)
AWBK31655 10 reactions
EUR 647
Description:
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GLI2 Western Blot kit (AWBK31885)
AWBK31885 10 reactions
EUR 647
Description:
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PAX4 Western Blot kit (AWBK32064)
AWBK32064 10 reactions
EUR 647
Description:
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  • Species reactivity:
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AHR Western Blot kit (AWBK32243)
AWBK32243 10 reactions
EUR 647
Description:
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  • Species reactivity:
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GLI1 Western Blot kit (AWBK32368)
AWBK32368 10 reactions
EUR 647
Description:
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  • Species reactivity:
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PAX7 Western Blot kit (AWBK32393)
AWBK32393 10 reactions
EUR 647
Description:
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  • Species reactivity:
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LHX6 Western Blot kit (AWBK32553)
AWBK32553 10 reactions
EUR 647
Description:
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  • Species reactivity:
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HIPK2 Western Blot kit (AWBK32586)
AWBK32586 10 reactions
EUR 647
Description:
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  • Species reactivity:
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PAX7 Western Blot kit (AWBK32742)
AWBK32742 10 reactions
EUR 647
Description:
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  • Species reactivity:
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ZFP67 Western Blot kit (AWBK32749)
AWBK32749 10 reactions
EUR 647
Description:
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  • Species reactivity:
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OLIG2 Western Blot kit (AWBK32753)
AWBK32753 10 reactions
EUR 647
Description:
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  • Species reactivity:
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ENG Western Blot kit (AWBK33068)
AWBK33068 10 reactions
EUR 647
Description:
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  • Species reactivity:
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SOX5 Western Blot kit (AWBK33323)
AWBK33323 10 reactions
EUR 647
Description:
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SOX10 Western Blot kit (AWBK33326)
AWBK33326 10 reactions
EUR 647
Description:
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RPS16 Western Blot kit (AWBK33535)
AWBK33535 10 reactions
EUR 647
Description:
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  • Species reactivity:
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ATP7A Western Blot kit (AWBK33797)
AWBK33797 10 reactions
EUR 647
Description:
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  • Species reactivity:
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A1BG Western Blot kit (AWBK33810)
AWBK33810 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity: Human
  • Application:
  • Assay info:
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NPM1 Western Blot kit (AWBK34094)
AWBK34094 10 reactions
EUR 647
Description:
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  • Species reactivity:
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LIG4 Western Blot kit (AWBK34122)
AWBK34122 10 reactions
EUR 647
Description:
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RNF2 Western Blot kit (AWBK34290)
AWBK34290 10 reactions
EUR 647
Description:
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  • Species reactivity:
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ZFP36 Western Blot kit (AWBK34385)
AWBK34385 10 reactions
EUR 647
Description:
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ARIH2 Western Blot kit (AWBK34418)
AWBK34418 10 reactions
EUR 647
Description:
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  • Species reactivity:
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KEAP1 Western Blot kit (AWBK34728)
AWBK34728 10 reactions
EUR 647
Description:
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  • Species reactivity:
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CLIC1 Western Blot kit (AWBK35045)
AWBK35045 10 reactions
EUR 647
Description:
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  • Species reactivity:
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KCNN2 Western Blot kit (AWBK35094)
AWBK35094 10 reactions
EUR 647
Description:
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  • Species reactivity:
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CLCN3 Western Blot kit (AWBK35504)
AWBK35504 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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  • Assay info:
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P2RX7 Western Blot kit (AWBK35518)
AWBK35518 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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TRPM3 Western Blot kit (AWBK35609)
AWBK35609 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
  • Application:
  • Assay info:
  • Sensitivity:

PITX2 Western Blot kit (AWBK35634)
AWBK35634 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
  • Application:
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ILF2 Western Blot kit (AWBK35731)
AWBK35731 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
  • Application:
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ONECUT2 Western Blot kit (AWBK35750)
AWBK35750 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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ZFP42 Western Blot kit (AWBK36244)
AWBK36244 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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ANXA1 Western Blot kit (AWBK36568)
AWBK36568 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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SPP1 Western Blot kit (AWBK36677)
AWBK36677 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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RUNX2 Western Blot kit (AWBK36678)
AWBK36678 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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ZBP1 Western Blot kit (AWBK36711)
AWBK36711 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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Bapx1 Western Blot kit (AWBK36826)
AWBK36826 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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G3BP Western Blot kit (AWBK37713)
AWBK37713 10 reactions
EUR 647
Description:
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  • Species reactivity:
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TWIST1 Western Blot kit (AWBK37997)
AWBK37997 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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ID4 Western Blot kit (AWBK38058)
AWBK38058 10 reactions
EUR 647
Description:
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  • Species reactivity:
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SATB1 Western Blot kit (AWBK38208)
AWBK38208 10 reactions
EUR 647
Description:
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  • Species reactivity:
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EED Western Blot kit (AWBK38384)
AWBK38384 10 reactions
EUR 647
Description:
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  • Species reactivity:
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TARDBP Western Blot kit (AWBK38941)
AWBK38941 10 reactions
EUR 647
Description:
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  • Species reactivity:
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TARDBP Western Blot kit (AWBK38942)
AWBK38942 10 reactions
EUR 647
Description:
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  • Species reactivity:
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BRD7 Western Blot kit (AWBK39018)
AWBK39018 10 reactions
EUR 647
Description:
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  • Species reactivity:
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JMJD3 Western Blot kit (AWBK40102)
AWBK40102 10 reactions
EUR 647
Description:
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  • Species reactivity:
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SNRP70 Western Blot kit (AWBK40439)
AWBK40439 10 reactions
EUR 647
Description:
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  • Species reactivity:
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SSB Western Blot kit (AWBK40462)
AWBK40462 10 reactions
EUR 647
Description:
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  • Species reactivity:
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STAU1 Western Blot kit (AWBK40537)
AWBK40537 10 reactions
EUR 647
Description:
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  • Species reactivity:
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PCBP1 Western Blot kit (AWBK40630)
AWBK40630 10 reactions
EUR 647
Description:
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  • Species reactivity:
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HNRPLL Western Blot kit (AWBK41102)
AWBK41102 10 reactions
EUR 647
Description:
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  • Species reactivity:
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MSI2 Western Blot kit (AWBK41112)
AWBK41112 10 reactions
EUR 647
Description:
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  • Species reactivity:
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HNRPA3 Western Blot kit (AWBK41195)
AWBK41195 10 reactions
EUR 647
Description:
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  • Species reactivity:
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WNT2B Western Blot kit (AWBK41254)
AWBK41254 10 reactions
EUR 647
Description:
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  • Species reactivity:
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GNB1L Western Blot kit (AWBK41328)
AWBK41328 10 reactions
EUR 647
Description:
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  • Species reactivity:
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SLC12A1 Western Blot kit (AWBK41388)
AWBK41388 10 reactions
EUR 647
Description:
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  • Species reactivity:
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TPM1 Western Blot kit (AWBK41392)
AWBK41392 10 reactions
EUR 647
Description:
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  • Species reactivity:
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CEACAM6 Western Blot kit (AWBK41504)
AWBK41504 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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SLC22A1 Western Blot kit (AWBK41516)
AWBK41516 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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STK11 Western Blot kit (AWBK41743)
AWBK41743 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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OTC Western Blot kit (AWBK41766)
AWBK41766 10 reactions
EUR 647
Description:
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  • Species reactivity:
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TSHR Western Blot kit (AWBK41870)
AWBK41870 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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RHOBTB1 Western Blot kit (AWBK41883)
AWBK41883 10 reactions
EUR 647
Description:
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  • Species reactivity:
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  • Assay info:
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KRT13 Western Blot kit (AWBK42031)
AWBK42031 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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  • Assay info:
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IDH3A Western Blot kit (AWBK42237)
AWBK42237 10 reactions
EUR 647
Description:
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  • Species reactivity:
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SLC1A5 Western Blot kit (AWBK42247)
AWBK42247 10 reactions
EUR 647
Description:
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  • Species reactivity:
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PBEF1 Western Blot kit (AWBK42255)
AWBK42255 10 reactions
EUR 647
Description:
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  • Species reactivity:
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FST Western Blot kit (AWBK42287)
AWBK42287 10 reactions
EUR 647
Description:
  • Description of target:
  • Species reactivity:
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SLC22A7 Western Blot kit (AWBK42306)
AWBK42306 10 reactions
EUR 647
Description:
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  • Species reactivity:
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ASPN Western Blot kit (AWBK42487)
AWBK42487 10 reactions
EUR 647
Description:
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  • Species reactivity:
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RSAD2 Western Blot kit (AWBK42696)
AWBK42696 10 reactions
EUR 647
Description:
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  • Species reactivity:
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PANX2 Western Blot kit (AWBK42778)
AWBK42778 10 reactions
EUR 647
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  • Species reactivity:
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Simple Western: Bringing the Western Blot 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 Western Blots

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 Western Blot 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.

 

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