p21 Cip1 Antibody | Affinity Biosciences

Product:	p21 Cip1 Antibody
Catalog:	AF6290
Description:	Rabbit polyclonal antibody to p21 Cip1
Application:	WB IF/ICC
Reactivity:	Human, Mouse, Rat
Prediction:	Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.:	21kDa; 18kD(Calculated).
Uniprot:	P38936
RRID:	AB_2827699

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

Size	Price	Inventory
100ul	$280	In stock
200ul	$350	In stock

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MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse,Rat

Prediction:

Bovine(100%), Horse(91%), Sheep(100%), Rabbit(83%), Dog(100%)

Clonality:

Polyclonal

Specificity:

p21 Cip1 Antibody detects endogenous levels of total p21 Cip1.

RRID:

AB_2827699
Cite Format: Affinity Biosciences Cat# AF6290, RRID:AB_2827699.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Storage:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Alias:

Fold/Unfold

CAP20; CDK-interacting protein 1; CDKI; CDKN1; Cdkn1a; CDN1A_HUMAN; CIP1; Cyclin Dependent Kinase Inhibitor 1A; Cyclin-dependent kinase inhibitor 1; Cyclin-dependent kinase inhibitor 1A (P21); Cyclin-dependent kinase inhibitor 1A (p21, Cip1); DNA Synthesis Inhibitor; MDA-6; MDA6; Melanoma differentiation-associated protein 6; Melanoma differentiation-associated protein; p21; P21 protein; p21CIP1; p21Cip1/Waf1; p21WAF; PIC1; SDI1; SLC12A9; WAF1; Wild type p53 activated fragment 1 (WAF1); Wild type p53 activated fragment 1; Wildtype p53-activated fragment 1;

Immunogens

Immunogen:

Uniprot:

P38936(CDN1A_HUMAN) >>Visit HPA database.

Gene(ID):

CDKN1A(1026)

Expression:

P38936 CDN1A_HUMAN:

Expressed in all adult tissues, with 5-fold lower levels observed in the brain.

Description:

This gene encodes a potent cyclin-dependent kinase inhibitor. The encoded protein binds to and inhibits the activity of cyclin-CDK2 or -CDK4 complexes, and thus functions as a regulator of cell cycle progression at G1. The expression of this gene is tightly controlled by the tumor suppressor protein p53, through which this protein mediates the p53-dependent cell cycle G1 phase arrest in response to a variety of stress stimuli.

Sequence:

P38936 CDN1A_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MSEPAGDVRQNPCGSKACRRLFGPVDSEQLSRDCDALMAGCIQEARERWNFDFVTETPLEGDFAWERVRGLGLPKLYLPTGPRRGRDELGGGRRPGTSPALLQGTAEEDHVDLSLSCTLVPRSGEQAEGSPGGPGDSQGRKRRQTSMTDFYHSKRRLIFSKRKP

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Bovine

100

Sheep

100

Dog

100

Horse

Rabbit

Zebrafish

Xenopus

Pig

Chicken

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P38936 As Substrate

P38936

Site	PTM Type	Enzyme	Source
S2	Acetylation		Uniprot
S15	Phosphorylation		Uniprot
K16	Ubiquitination		Uniprot
S31	Phosphorylation		Uniprot
T57	Phosphorylation	Q16539 (MAPK14) , P28482 (MAPK1) , P45983 (MAPK8) , P49841 (GSK3B)	Uniprot
K75	Ubiquitination		Uniprot
T80	Phosphorylation		Uniprot
S98	Phosphorylation	Q99683 (MAP3K5) , P45983 (MAPK8)	Uniprot
S114	Phosphorylation	P49841 (GSK3B)	Uniprot
S116	Phosphorylation		Uniprot
S123	Phosphorylation		Uniprot
S130	Phosphorylation	Q16539 (MAPK14) , P45983 (MAPK8) , P24941 (CDK2) , P28482 (MAPK1) , Q00534 (CDK6)	Uniprot
K141	Acetylation		Uniprot
K141	Ubiquitination		Uniprot
T145	Phosphorylation	P31749 (AKT1) , Q9P1W9 (PIM2) , O14757 (CHEK1) , O43293 (DAPK3) , P11309 (PIM1)	Uniprot
S146	Phosphorylation	P53355 (DAPK1) , O14757 (CHEK1) , P11309 (PIM1) , Q9NRM7 (LATS2) , Q9P1W9 (PIM2) , P17252 (PRKCA) , Q05655 (PRKCD) , P31749 (AKT1) , Q15208 (STK38)	Uniprot
Y151	Phosphorylation		Uniprot
S153	Phosphorylation	P17252 (PRKCA) , Q9Y463 (DYRK1B)	Uniprot
K154	Acetylation		Uniprot
K154	Ubiquitination		Uniprot
R156	Methylation		Uniprot
S160	Phosphorylation		Uniprot
K161	Acetylation		Uniprot
K161	Ubiquitination		Uniprot
K163	Acetylation		Uniprot
K163	Ubiquitination		Uniprot

Research Backgrounds

P38936_CDN1A_HUMAN

Function:

May be involved in p53/TP53 mediated inhibition of cellular proliferation in response to DNA damage. Binds to and inhibits cyclin-dependent kinase activity, preventing phosphorylation of critical cyclin-dependent kinase substrates and blocking cell cycle progression. Functions in the nuclear localization and assembly of cyclin D-CDK4 complex and promotes its kinase activity towards RB1. At higher stoichiometric ratios, inhibits the kinase activity of the cyclin D-CDK4 complex. Inhibits DNA synthesis by DNA polymerase delta by competing with POLD3 for PCNA binding. Plays an important role in controlling cell cycle progression and DNA damage-induced G2 arrest.

PTMs:

Phosphorylation of Thr-145 by Akt or of Ser-146 by PKC impairs binding to PCNA. Phosphorylation at Ser-114 by GSK3-beta enhances ubiquitination by the DCX(DTL) complex. Phosphorylation of Thr-145 by PIM2 enhances CDKN1A stability and inhibits cell proliferation. Phosphorylation of Thr-145 by PIM1 results in the relocation of CDKN1A to the cytoplasm and enhanced CDKN1A protein stability. UV radiation-induced phosphorylation at Thr-80 by LKB1 and at Ser-146 by NUAK1 leads to its degradation.

Ubiquitinated by MKRN1; leading to polyubiquitination and 26S proteasome-dependent degradation. Ubiquitinated by the DCX(DTL) complex, also named CRL4(CDT2) complex, leading to its degradation during S phase or following UV irradiation. Ubiquitination by the DCX(DTL) complex is essential to control replication licensing and is PCNA-dependent: interacts with PCNA via its PIP-box, while the presence of the containing the 'K+4' motif in the PIP box, recruit the DCX(DTL) complex, leading to its degradation. Ubiquitination at Ser-2 leads to degradation by the proteasome pathway. Ubiquitinated by RNF114; leading to proteasomal degradation.

Acetylation leads to protein stability. Acetylated in vitro on Lys-141, Lys-154, Lys-161 and Lys-163. Deacetylation by HDAC1 is prevented by competitive binding of C10orf90/FATS to HDAC1 (By similarity).

Subcellular Location:

Cytoplasm. Nucleus.

Tissue Specificity:

Expressed in all adult tissues, with 5-fold lower levels observed in the brain.

Subunit Structure:

Interacts with HDAC1; the interaction is prevented by competitive binding of C10orf90/FATS to HDAC1 facilitating acetylation and protein stabilization of CDKN1A/p21 (By similarity). Interacts with MKRN1. Interacts with PSMA3. Interacts with PCNA. Component of the ternary complex, cyclin D-CDK4-CDKN1A. Interacts (via its N-terminal domain) with CDK4; the interaction promotes the assembly of the cyclin D-CDK4 complex, its nuclear translocation and promotes the cyclin D-dependent enzyme activity of CDK4. Binding to CDK2 leads to CDK2/cyclin E inactivation at the G1-S phase DNA damage checkpoint, thereby arresting cells at the G1-S transition during DNA repair. Interacts with PIM1. Interacts with STK11 and NUAK1. Interacts wih DTL. Interacts with isoform 1 and isoform 2 of TRIM39.

Family&Domains:

The PIP-box K+4 motif mediates both the interaction with PCNA and the recruitment of the DCX(DTL) complex: while the PIP-box interacts with PCNA, the presence of the K+4 submotif, recruits the DCX(DTL) complex, leading to its ubiquitination.

The C-terminal is required for nuclear localization of the cyclin D-CDK4 complex.

Belongs to the CDI family.

Research Fields

· Cellular Processes > Cell growth and death > Cell cycle. (View pathway)

· Cellular Processes > Cell growth and death > p53 signaling pathway. (View pathway)

· Cellular Processes > Cell growth and death > Cellular senescence. (View pathway)

· Environmental Information Processing > Signal transduction > ErbB signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > FoxO signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > Jak-STAT signaling pathway. (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > HTLV-I infection.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer. (View pathway)

· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Colorectal cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Renal cell carcinoma. (View pathway)

· Human Diseases > Cancers: Specific types > Pancreatic cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Endometrial cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Glioma. (View pathway)

· Human Diseases > Cancers: Specific types > Prostate cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Thyroid cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Basal cell carcinoma. (View pathway)

· Human Diseases > Cancers: Specific types > Melanoma. (View pathway)

· Human Diseases > Cancers: Specific types > Bladder cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Chronic myeloid leukemia. (View pathway)

· Human Diseases > Cancers: Specific types > Small cell lung cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Non-small cell lung cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Breast cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma. (View pathway)

· Human Diseases > Cancers: Specific types > Gastric cancer. (View pathway)

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

References

1). Sirt3-mediated mitophagy regulates AGEs-induced BMSCs senescence and senile osteoporosis. Redox Biology, 2021 (PubMed: 33662874) [IF=11.4]

Application: WB Species: mice Sample: bone marrow mesenchymal stem (BMSCs)

Fig. 1. Effects of AGEs in different concentrations on the senescence of BMSCs. The BMSCs were treated with AGEs (50–200 μg/mL) or BSA for 24–72 h. (A) SA-β-gal assay for detection of BMSCs senescence. Scale bar: 100 μm. (B) Detection of H3K9me3 by immunofluorescence in BMSCs. Scale bar: 100 μm. (C) Detection of γ-H2AX by immunofluorescence in BMSCs. Scale bar: 100 μm. (D–I) Representative Western blotting assay and quantitation of the level of P16, P21, P53. **p < 0.01 versus BSA.

2). KLF12 promotes the proliferation of breast cancer cells by reducing the transcription of p21 in a p53-dependent and p53-independent manner. Cell Death & Disease, 2023 (PubMed: 37156774) [IF=9.0]

Application: WB Species: Human Sample: MCF7 cells

Fig. 4 KLF12 reduces the stability of p53. a Western blot assay showed changes of exogenous p53 in HEK293T cells transfected with increasing doses of KLF12. b Changes in endogenous p53 and p21 in MCF7 cells transfected with increasing doses of KLF12 by Western blot assay. c Changes in the endogenous level of p53 and p21 in ZR-75-30 cells transfected with NC, shKLF12#1, or shKLF12#2 by Western blot assay. d Changes in exogenous p53 in HEK293T cells transfected with GFP-KLF12 without and with 20 μg/ml MG132 treatment by Western blot assay. e, f The half-life of p53 in MCF-7 cells with overexpressed KLF12 treated with CHX (50 μg/ml) by Western blot assay. g CoIP assay showed changes in the level of ubiquitinated p53 in HEK293T cells transfected with Flag-KLF12 plus 4 h treatment with 20 μg/ml MG132. h CoIP assay detected changes in K48-linked polyubiquitylation of p53 and K63-linked polyubiquitylation of p53 in HEK293T cells. The cells were treated with MG132 for 4 h following transfection.

3). Histone acetylation plays an important role in MC-LR-induced apoptosis and cycle disorder in SD rat testicular cells. Chemosphere, 2020 (PubMed: 31683423) [IF=8.8]

Application: WB Species: Rat Sample: testicular tissue

Fig4. The effect of TSA and MC-LR on cell cycle-related genes and proteins 434 levels in co-cultured Sertoli-germ cells and SD rat testicular tissues. (A) 435 Expressions of cell cycle-related genes in vitro and (B) expressions of cell 436 cycle-related genes in vivo were detected by RT-qPCR. (C) Expressions of cell cycle-related proteins in vitro and (D) expressions of cell cycle-related proteins in 438 vivo were detected by western blotting. (E) Quantitative analysis of proteins 439 expression levels in vitro. (F) Quantitative analysis of proteins expression levels in 440 vivo. (*P<0.05 vs. the control group; #P<0.05 vs. the 36 µM MC-LR group or the 40 441 µg kg-1 MC-LR group)

4). d-Borneol enhances cisplatin sensitivity via p21/p27-mediated S-phase arrest and cell apoptosis in non-small cell lung cancer cells and a murine xenograft model. Cellular & Molecular Biology Letters, 2022 (PubMed: 35883026) [IF=8.3]

5). Downregulation of VEGFA accelerates AGEs-mediated nucleus pulposus degeneration through inhibiting protective mitophagy in high glucose environments. International journal of biological macromolecules, 2024 (PubMed: 38320636) [IF=8.2]

6). Muscone restores anoikis sensitivity in TMZ-resistant glioblastoma cells by suppressing TOP2A via the EGFR/Integrin β1/FAK signaling pathway. Phytomedicine : international journal of phytotherapy and phytopharmacology, 2024 (PubMed: 38723526) [IF=7.9]

7). Amelioratory effects of astragaloside IV on hepatocarcinogenesis via Nrf2-mediated pSmad3C/3L transformation. Phytomedicine, 2023 (PubMed: 37301185) [IF=7.9]

8). VDR activation attenuates osteoblastic ferroptosis and senescence by stimulating the Nrf2/GPX4 pathway in age-related osteoporosis. Free Radical Biology and Medicine, 2022 (PubMed: 36402439) [IF=7.4]

9). Integrating scRNA and bulk-RNA sequencing develops a cell senescence signature for analyzing tumor heterogeneity in clear cell renal cell carcinoma. Frontiers in Immunology, 2023 (PubMed: 37503331) [IF=7.3]

Application: WB Species: Human Sample: 786-O cell

Figure 8 Construction of risk model and functional validation of critical genes. (A) Differential analysis between clust1 and no_clust1 in TCGA dataset; (B) Differential analysis between clust2 and no_clust2 in TCGA dataset; (C) Differential analysis between clust3 and no_clust3 in TCGA dataset; (D) A total of 961 promising candidates were identified among the differentially expressed genes; (E) Trajectory of each independent variable as lambda changes; (F) Confidence interval under lambda; (G) Coefficients of prognostic-related genes obtained from multivariate Cox analysis; (H) Western blot assay of DUSP1 protein expression in 786-O cell after transfection of Si-DUSP1; (I) Colony formation assay was carried out to evaluate the proliferation of 786-O cell; (J, K) Transwell assay was used to assess the migration and invasion of 786-O cell. (L) Western blot assay of p21 and p53 protein expression in 786-O cell after transfection of Si-DUSP1. *, p< 0.05; (M) SA-β-gal staining of 786-O cell. *, p< 0.05.

10). Primary microcephaly gene CENPE is a novel biomarker and potential therapeutic target for non-WNT/non-SHH medulloblastoma. Frontiers in Immunology, 2023 (PubMed: 37593739) [IF=7.3]

Application: WB Species: Human Sample:

Figure 9 CENPE regulates the cell cycle and p53 pathway of non-WNT/non-SHH MB. (A–E) Western blot showed that the protein level of CENPE, P53, P21 and CDK1 by the knockdown of CENPE (mean ± SD, n = 3). (F–H) Flow cytometry was used to detect the cell cycle (mean ± SD, n = 3).

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p21 Cip1 Antibody - #AF6290