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Recombinant fragment corresponding to Human CDKN2A/p16INK4a .
Database link: Q8N726
Our Abpromise guarantee covers the use of ab54210 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/2000. Tested using a recombinant protein expressed in E. coli. We have no data on the detection of the endogenous protein and would welcome feedback from anyone that has performed a WB with ab54210 using cell lysates.|
|ICC/IF||Use at an assay dependent concentration. PubMed: 24828678|
|IHC-P||1/200 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Recommended antigen retrieval: Place slides in a special glass slide holder and fill in the rack with slides to ensure even heating. Rinse rack in 1L of Citrate buffer in ordinary pressure cooker,(0.01M,pH6.0) to immerse all slides. Cover the cap and heat for 15-20 minutes on electric cooker until water boils. Begin timing, only 2 minutes in high-pressure Remove the pressure cooker, Allow it to cool(Cool the slides at room temperature) Rinse the slides in PBS for 5 minutes|
Overlay histogram showing HeLa cells stained with ab54210 (red line). The cells were fixed with 80% methanol (5 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 (ab54210, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.
ab54210 (1:500) staining p16 INK4a in rat liver tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (by DAB staining) is shown.
ab54210 (1:500) staining p16 INK4a in human brain tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (by DAB staining) is shown.
ab54210 (1:500) staining p16 INK4a in human brain tumor tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (by DAB staining) is shown.
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