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Anti-CDKN2A/p16INK4a Rabbit Antibody [B12M16]

Catalog No.: F3773

Application: Reactivity: Human

Experiment Essentials

WB
Recommended wet transfer conditions: 200 mA, 60 min，Recommended to use 0.22 μm PVDF membrane.

Usage Information

Dilution
WB1:1000 IP1:30 IHC1:500 IF1:50 FCM1:500

Application
WB, IP, IHC, IF , FCM

Reactivity
Human

Source
Rabbit

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
17 kDa 17 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight.

Positive Control	Human cervical cancer tissue; Human endometrial cancer tissue; HEK-293T; HeLa; HEK-293; Saos-2
Negative Control	Human cervix tissue; MCF7; MDA-MB-231

Datasheet & SDS

Biological Description

Specificity
CDKN2A/p16INK4a (B12M16) Rabbit mAb detects endogenous levels of total CDKN2A/p16INK4a protein.

Clone
B12M16

Synonym(s)
CDKN2, MTS1, CDKN2A, Cyclin-dependent kinase inhibitor 2A, Cyclin-dependent kinase 4 inhibitor A, Multiple tumor suppressor 1, p16-INK4a, CDK4I, MTS-1, p16-INK4, p16INK4A

Background
CDKN2A encodes p16INK4a, a key tumor suppressor protein belonging to the INK4 family, characterized by four ankyrin repeats critical for binding cyclin-dependent kinases 4 and 6 (CDK4/6). The primary function of p16INK4a is to inhibit CDK4/6 activity, preventing phosphorylation of the retinoblastoma protein (Rb), thereby halting cell cycle progression from the G1 to S phase through blocking the release of E2F transcription factors. This regulation is essential for controlling cell proliferation and preventing uncontrolled cell division. Expression of p16INK4a is tightly controlled at multiple levels, including transcriptional, epigenetic, and post-translational mechanisms. In particular, SIRT7-mediated deacetylation of histone markers represses CDKN2A transcription in non-senescent cells, whereas loss of repression leads to increased p16INK4a expression and induction of cell cycle arrest during senescence. p16INK4a also contributes significantly to cellular senescence by promoting irreversible growth arrest in response to cellular stress and aging. Mutations, deletions, or promoter methylation of CDKN2A disrupt p16INK4a function and contribute to tumorigenesis.

Background

CDKN2A encodes p16INK4a, a key tumor suppressor protein belonging to the INK4 family, characterized by four ankyrin repeats critical for binding cyclin-dependent kinases 4 and 6 (CDK4/6). The primary function of p16INK4a is to inhibit CDK4/6 activity, preventing phosphorylation of the retinoblastoma protein (Rb), thereby halting cell cycle progression from the G1 to S phase through blocking the release of E2F transcription factors. This regulation is essential for controlling cell proliferation and preventing uncontrolled cell division. Expression of p16INK4a is tightly controlled at multiple levels, including transcriptional, epigenetic, and post-translational mechanisms. In particular, SIRT7-mediated deacetylation of histone markers represses CDKN2A transcription in non-senescent cells, whereas loss of repression leads to increased p16INK4a expression and induction of cell cycle arrest during senescence. p16INK4a also contributes significantly to cellular senescence by promoting irreversible growth arrest in response to cellular stress and aging. Mutations, deletions, or promoter methylation of CDKN2A disrupt p16INK4a function and contribute to tumorigenesis.

References
https://pubmed.ncbi.nlm.nih.gov/21619050/ https://pubmed.ncbi.nlm.nih.gov/30482142/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%