Anti-p15 INK4b antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p15 INK4b antibody (AB53034)

ab53034 (2µg/ml) staining p15 INK4b in human colon carcinoma.
Sections were stained using an automated system (DAKO Autostainer Plus ), at room temperature : sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0 . Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-p15 INK4b antibody (AB53034)

Immunofluorescence analysis of HeLa cells, using Anti-p15 INK4b antibody (ab53034). The picture on the right is blocked with the synthesized peptide.

• WB

Unknown

Western blot - Anti-p15 INK4b antibody (AB53034)

All lanes:

Western blot - Anti-p15 INK4b antibody (ab53034) at 1/500 dilution

Lane 1:

HeLa cell extract

Lane 2:

HeLa cell extract with peptide

Predicted band size: 15 kDa

Observed band size: 15 kDa

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• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p15 INK4b antibody (AB53034)

Left panel : with primary antibody at 2 ug/ml. Right panel : isotype control.
Sections were stained using an automated system (Dako PT Link), at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffers EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

• WB

CiteAb

Western blot - Anti-p15 INK4b antibody (AB53034)

p15 INK4b western blot using anti-p15 INK4b antibody ab53034. Publication image and figure legend from Tanaka, T., Terai, Y., et al., 2017, PLoS One, PubMed 28472136.

ab53034 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab53034 please see the product overview.

(A) A Western blot analysis shows the expression of Rb and phospho-Rb protein in HEC1A and HEC108 cells. Both cell lines demonstrated expression of cyclin D1. Strong expression of p15, p16, p18 and p19, which act as inhibitors of CDK 4/6, was not seen. (B) Effect of PD-0332991 on phosphorylation of Rb protein. HEC1A (top) and HEC108 (bottom) cells showed a time- and concentration-dependent inhibition of Rb phosphorylation following treatment with a PD-0332991 inhibitor. However, PD-0332991 inhibited total Rb expression to a greater extent than phospho-Rb.

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