Product nameAnti-Hsp70 antibody [N27F3-4]
See all Hsp70 primary antibodies
DescriptionMouse monoclonal [N27F3-4] to Hsp70
SpecificityDetects 72 and 73kDa proteins corresponding to the molecular mass of inducible Hsp and Hsc70 of SDS PAGE immunoblots.
Tested applicationsSuitable for: IHC-Fr, IHC-P, WB, IP, Flow Cyt, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Sheep, Rabbit, Chicken, Guinea pig, Hamster, Cow, Dog, Human, Pig, Xenopus laevis, Caenorhabditis elegans, Carp, Drosophila melanogaster, Fish, Monkey, Apteronotus leptorhynchus
Predicted to work with: Plants
Recombinant hsp70/hsc70 (Human)
- HeLa nuclear extract lysate (ab14655) can be used as a positive control in WB. ICC/IF: Hek293 cells.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferPreservative: 0.09% Sodium Azide
Constituents: 50% Glycerol, PBS, pH 7.2
Concentration information loading...
PurityProtein G purified
Our Abpromise guarantee covers the use of ab47454 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 70 kDa.|
|IP||Use a concentration of 5 µg/ml.|
|Flow Cyt||Use at an assay dependent concentration.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|ICC/IF||Use a concentration of 5 µg/ml.|
RelevanceFunction: In cooperation with other chaperones, the Hsp70 family 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. Tissue specificity: HSPA1B is testis-specific.
Cellular localizationCytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
- DnaK type molecular chaperone HSP70 1 antibody
- Epididymis secretory protein Li 103 antibody
- FLJ54303 antibody
Hsp70 was immunoprecipitated using 0.5mg Rat Testis tissue lysate, 5µg of Mouse monoclonal to Hsp70 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Rat Testis tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab47454.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.
Band: 70kDa, non specific bands - 50kDa: We are unsure as to the identity of this extra band; Hsp70
Immunohistochemistry analysis using ab47454 (1:100) staining paraffin-embedded mouse backskin. Stained for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT.
Anti-Hsp70 antibody [N27F3-4] (ab47454) at 1/500 dilution + Apteronotus leptorhynchus brain tissue lysate at 50 µg
Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal at 1/1000 dilution
Performed under reducing conditions.
Predicted band size: 70 kDa
Observed band size: 70 kDa
Exposure time: 5 minutes
All lanes : Anti-Hsp70 antibody [N27F3-4] (ab47454) at 1 µg/ml
Lane 1 : Cell lysate prepared from human A431 cell line
Lane 2 : Cell lysate prepared from human A549 cell line
Lane 3 : Cell lysate prepared from human HCT116 cell line
Lane 4 : Cell lysate prepared from human Hela cell line
Lane 5 : Cell lysate prepared from human HEK293 cell line
Lane 6 : Cell lysate prepared from human HepG2 cell line
Lane 7 : Cell lysate prepared from human HL-60 cell line
Lane 8 : Cell lysate prepared from human HUVEC cell line
Lane 9 : Cell lysate prepared from human Jurkat cell line
Lane 10 : Cell lysate prepared from human MCF7 cell line
Lane 11 : Cell lysate prepared from human PC3 cell line
Lane 12 : Cell lysate prepared from human T98G cell line
Lane 13 : Lysate prepared from rat brain tissue
Lysates/proteins at 10 µg per lane.
Predicted band size: 70 kDa
ICC/IF image of ab47454 stained Hek293 cells. The cells were 100% methanol fixed (5 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 (ab47454, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 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.