Primary antibodies against HSP90 (C45G5) and β-actin were purchased from Cell Signaling Technology. LC3B antibody was from GenTex. HSC70 (13D3) was purchased from Abcam. The anti-p53 antibody (DO-1) was a gift from Dr Xirodimas. Cell extracts were lysed in 2x SDS sample buffer. Proteins were resolved by SDS-PAGE and transferred to nitrocellulose or PVDF membranes using the Trans-Blot® Turbo™ Transfer System ( Bio-Rad). Peroxidase-coupled anti-mouse and anti-rabbit secondary antibodies were used at a dilution of 1:10.000 (Sigma). Bound antibodies were detected by enhanced chemiluminescence (Millipore). To obtain the image for the western blotting the Molecular Imager Gel Doc XRS system (Biorad) was used, this system provides a reliable and sensitive imaging of chemilunescence western blots. For the quantification and analysis, we used the Image Lab Software (Biorad), a powerful and specific software for acquisition, analysis and quantification of blot images.
Hsc70 13d3
Manufactured by Abcam
HSC70 (13D3) is a monoclonal antibody that recognizes the Heat Shock Cognate 71 kDa Protein (HSC70). HSC70 is a constitutively expressed member of the heat shock protein 70 (HSP70) family and plays a crucial role in the regulation of protein folding and assembly.
Automatically generated - may contain errors
Lab products found in correlation
Enhanced chemiluminescence, by Merck Group (1 mentions)
Molecular imager gel doc xrs system, by Bio-Rad (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Image lab software, by Bio-Rad (1 mentions)
Trans blot turbo transfer system, by Bio-Rad (1 mentions)
β actin c4, by Santa Cruz Biotechnology (1 mentions)
Image lab 5, by Bio-Rad (1 mentions)
Bradford protein assay, by Bio-Rad (1 mentions)
Chemidoc xrs system, by Bio-Rad (1 mentions)
2 protocols using hsc70 13d3
1
Western Blot Analysis of HSP90, p53, and Actin
2
Monocyte Protein Expression Analysis
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Monocytes were lysed in RIPA buffer containing a cocktail of protease inhibitors. Protein content was determined using Bradford Protein Assay (Bio-Rad). Protein lysates were separated on 8% SDS-PAGE and transferred on polyvinylidene uoride membrane. Membranes were probed overnight with the following monoclonal antibodies: Plin2 AP125 (LifeSpanBioSciences,Inc), LAMP2A H4B4 (Abcam), Hsc70 13D3 (Abcam) and β-actin C-4 (Santa Cruz). Detection and analysis were performed with the ChemiDoc XRS system with Image Lab 5.0 software (Bio-Rad). All results were normalized with β-actin and expressed as ratio.
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