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|Sample type||Average %||Range|
|Cell culture extracts||105||104% - 106%|
Hsp27 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of mouse Hsp27 protein in cell extracts.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
Heat shock proteins (HSPs) are molecular chaperones that facilitate the proper folding and function of proteins within the cell. Heat shock protein 27 (Hsp27), also known as heat shock protein beta-1 (HspB1) is a 209 amino acid protein encoded by the HSPB1 gene. Hsp27 a member of the sHsp (small heat shock protein) group, whose functions are chaperone activity, thermotolerance, inhibition of apoptosis, and regulation of cellular development and differentiation. Hsp70 member proteins may assist inhibition of apoptosis by acting on the caspase-dependent pathway and against agents such as tumor necrosis factor-alpha, staurosporine, and doxorubicin. Hsp27 can form oligomers depending on the phosphorylation state of the protein and during heat shock. The oligomers are connected to the chaperone activity, while dimers have limited activity. Hsp27 interacts with actin and intermediate filaments, which provides protection to actin filaments from fragmentation.
|Components||1 x 96 tests|
|10X Mouse Hsp27 Detector Antibody||1 x 600µl|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|50X Cell Extraction Enhancer Solution (ab193971)||1 x 1ml|
|5X Cell Extraction Buffer PTR (ab193970)||1 x 10ml|
|Antibody Diluent CPR||1 x 6ml|
|Mouse Hsp27 Capture Antibody Lyophilized||1 vial|
|Mouse Hsp27 Lyophilized Recombinant Protein||2 vials|
|Plate Seals||1 unit|
|Sample Diluent NS||1 x 12ml|
|SimpleStep Pre-Coated 96-Well Microplate (ab206978)||1 unit|
|Stop Solution||1 x 12ml|
|TMB Development Solution||1 x 12ml|
Our Abpromise guarantee covers the use of ab216949 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||Use at an assay dependent concentration.|
Background-subtracted data values (mean +/- SD) are graphed.
The concentrations of Hsp27 were measured in duplicate and interpolated from the Hsp27 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Hsp27 concentration was determined to be 540 ng/mg in C2C12 cell extract.
The concentrations of Hsp27 were measured in duplicate and interpolated from the Hsp27 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Hsp27 concentrations were determined to be 680 ng/mg in mock-treated, 1470 ng/mg in heat-shocked, and 700 ng/mg in untreated C2C12 cell extract.
ab216949 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"