The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 - 5 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 73 kDa (predicted molecular weight: 70 kDa).
In cooperation with other chaperones, Hsp70s stabilize preexistent proteins against aggregation and mediate the folding of newly translated polypeptides in the cytosol as well as within organelles. These chaperones participate in all these processes through their ability to recognize nonnative conformations of other proteins. They bind extended peptide segments with a net hydrophobic character exposed by polypeptides during translation and membrane translocation, or following stress-induced damage. In case of rotavirus A infection, serves as a post-attachment receptor for the virus to facilitate entry into the cell.
HSPA1B is testis-specific.
Belongs to the heat shock protein 70 family.
Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
ICC/IF image of ab94368 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab94368 at 5µg/ml overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti- rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in formaldehyde (4%, 10 minutes) fixed HeLa, and MCF-7 cells at 5ug/ml, and also in Methanol (100%, 5minutes) fixed HeLa, Hek293, HepG2, and MCF-7 cells at 1ug/ml.
Western blot - Anti-Hsp70 antibody (ab94368)
All lanes : Anti-Hsp70 antibody (ab94368) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 2 : Liver (Mouse) Tissue Lysate Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate Lane 4 : Colon (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Takata T et al. Generation of glyceraldehyde-derived advanced glycation end-products in pancreatic cancer cells and the potential of tumor promotion. World J Gastroenterol23:4910-4919 (2017).
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