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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Hsp70 aa 600-700 (C terminal).
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
A 40 µl trial size is available to purchase for this antibody.
Our Abpromise guarantee covers the use of ab45133 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/20000. Detects a band of approximately 85 kDa.|
|IHC-P||1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).
Overlay histogram showing HeLa cells fixed in 2% PFA and stained with purified ab45133 at a dilution of 1 in 20 (pink line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 150. Rabbit monoclonal IgG was used as an isotype control.
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunohistochemical staining of paraffin embedded human breast carcinoma with purified ab45133 at a working dilution of 1 in 400. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunofluorescence staining of A431 cells with purified ab45133 at a working dilution of 1 in 400, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 555 goat anti rabbit, used at a dilution of 1 in 500. The cells were fixed in 4% PFA. The negative control is shown in bottom right hand panel - for the negative control, purified ab45133 was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-mouse antibody (ab150113) at a dilution of 1/500.
Overlay histogram showing Jurkat cells stained with unpurified ab45133 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab45133, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Overlay histogram showing HeLa cells fixed in 2% PFA and stained with unpurified ab45133 at a dilution of 1 in 20 (pink line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 150. Rabbit monoclonal IgG was used as an isotype control.
Unpurified ab45133 at a 1/100 dilution staining human Hsp70 in human prostate carcinoma by immunohistochemistry using paraffin embedded tissue.Ab45133 at a 1/100 dilution staining human Hsp70 in human prostate carcinoma by immunohistochemistry using paraffin embedded tissue.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"