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

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.
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.
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:
Gene(ID):
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:
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
91
Rabbit
83
Zebrafish
67
Xenopus
56
Pig
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P38936 As Substrate

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

Research Backgrounds

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.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
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). Guo Y et al. Sirt3-mediated mitophagy regulates AGEs-induced BMSCs senescence and senile osteoporosis. Redox Biol 2021 May;41:101915. (PubMed: 33662874) [IF=9.986]

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). Wang Y et al. Histone acetylation plays an important role in MC-LR-induced apoptosis and cycle disorder in SD rat testicular cells. Chemosphere 2019 Oct 12;241:125073 (PubMed: 31683423) [IF=5.778]

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)

3). He P et al. AMPK Activity Contributes to G2 Arrest and DNA Damage Decrease via p53/p21 Pathways in Oxidatively Damaged Mouse Zygotes. Front Cell Dev Biol 2020 Sep 8;8:539485. (PubMed: 33015052) [IF=5.201]

Application: WB    Species: mouse    Sample: zygotes

FIGURE 5 | Activated AMPK regulates the oxidative stress induction of cell cycle regulatory proteins of p53 and p21 in mouse zygotes. CC, Compound C; SBI, SBI-0206965. (A) Representative data showing the level of p53, p21, p53-pSer15, p53-pSer20 and p53-pSer46 in control, H2O2-treated, Compound C/H2O2-treated, and SBI-0206965/H2O2-treated zygotes in the G2 phase. (B) Quantification data of figure (n = 3). (C) Representative image of the subcellular distribution and expression of p21 (green) using immunostaining. Nuclei were stained with DAPI (blue). Scale bar = 20 µm. (D) Mean fluorescence intensity of nuclear and cytoplasmic p21 in mouse zygotes in the G2 phase. N = 37 zygotes per group. Results in (B,D) were analyzed by one-way ANOVA and Tukey’s test. *p < 0.05; ***p < 0.001. Error bars: SD.

4). Yang H et al. Navitoclax (ABT263) reduces inflammation and promotes chondrogenic phenotype by clearing senescent osteoarthritic chondrocytes in osteoarthritis. Aging (Albany NY) 2020 Jul 1;12. (PubMed: 32611834) [IF=4.831]

5). Yao MZ et al. MEIS1 regulated proliferation and migration of pulmonary artery smooth muscle cells in hypoxia-induced pulmonary hypertension. Life Sci 2020 May 22;117822. (PubMed: 32450174) [IF=3.647]

6). Ji S et al. Resveratrol promotes oxidative stress to drive DLC1 mediated cellular senescence in cancer cells. Exp Cell Res 2018 Sep 15;370(2):292-302 (PubMed: 29964052) [IF=3.383]

7). Wang D et al. GRIM-19 inhibits proliferation and induces apoptosis in a p53-dependent manner in colorectal cancer cells through the SIRT7/PCAF/MDM2 axis. Exp Cell Res 2021 Oct 1;407(1):112799. (PubMed: 34461110) [IF=3.383]

8). Sun H et al. GCNT4 is Associated with Prognosis and Suppress Cell Proliferation in Gastric Cancer. Onco Targets Ther 2020 Aug 25;13:8601-8613. (PubMed: 32922038) [IF=3.337]

9). Sun Z et al. Metformin inhibits pulmonary artery smooth muscle cell proliferation by upregulating p21 via NONRATT015587. 2. Int J Mol Med 2022 Apr;49(4):49. (PubMed: 35147202) [IF=3.098]

10). Yuan Y et al. STAT3 stimulates adipogenic stem cell proliferation and cooperates with HMGA2 during the early stage of differentiation to promote adipogenesis. Biochem Biophys Res Commun 2017 Jan 22;482(4):1360-1366 (PubMed: 27940362)

Application: WB    Species: mouse    Sample:

STAT3 knockdown blocked cell cycle procession at S phase in adipogenic stem cells and early differentiating cells. (A) Flow cytometric analysis of 3T3-L1 stem cells at 48 h after siRNA introduction. Expression of cell cycle-associated genes was measured at mRNA (B) and protein (C) levels at 48 h after siRNA introduction in 3T3-L1 stem cells. (D) Flow cytometric analysis of 3T3-L1 cells that were differentiated for 24 h, with differentiation induced at 72 h after STAT3 siRNA introduction. qRT-PCR analysis (E) and Western blot analyses (F) of cell cycle-associated genes after STAT3 siRNA introduction. Values are presented as means ± S.D. of three replicates. * P < 0.01, # P < 0.05 vs siGL3.

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