Product nameAnti-p15 INK4b antibody
See all p15 INK4b primary antibodies
DescriptionRabbit polyclonal to p15 INK4b
SpecificityDetects endogenous levels of total p15 INK protein
Tested applicationsSuitable for: IP, WB, ELISA, IHC-P, ICC/IFmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat
Synthetic peptide derived from human p15 INK
- HeLa cell extract
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 50% Glycerol, 0.87% Sodium chloride, PBS
Without Mg+2 and Ca+2
Concentration information loading...
PurityImmunogen affinity purified
Purification notesab53034 was purified from rabbit antiserum by affinity chromatography using epitope specific immunogen
Our Abpromise guarantee covers the use of ab53034 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use a concentration of 5 µg/ml.|
|WB||1/500 - 1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).|
|IHC-P||Use a concentration of 2 µg/ml.|
|ICC/IF||Use at an assay dependent concentration.|
FunctionInteracts strongly with CDK4 and CDK6. Potent inhibitor. Potential effector of TGF-beta induced cell cycle arrest.
Sequence similaritiesBelongs to the CDKN2 cyclin-dependent kinase inhibitor family.
Contains 4 ANK repeats.
- Information by UniProt
- CDK inhibitory protein antibody
- CDK4B Inhibitor antibody
- Cdkn2b antibody
All lanes : 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
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.
Immunofluorescence analysis of HeLa cells, using Anti-p15 INK4b antibody (ab53034). The picture on the right is blocked with the synthesized peptide.
p15 INK4b was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to p15 INK4b 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, Hela whole cell extract 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 ab53034.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 15kDa; p15 INK4b
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.
This product has been referenced in:
- Gao S et al. Long noncoding RNA HOTAIR promotes the self-renewal of leukemia stem cells through epigenetic silencing of p15. Exp Hematol 67:32-40.e3 (2018). Read more (PubMed: 30172749) »
- Castex J et al. Inactivation of Lsd1 triggers senescence in trophoblast stem cells by induction of Sirt4. Cell Death Dis 8:e2631 (2017). Read more (PubMed: 28230862) »