p21 Cip1 Antibody - #DF6423

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Product: p21 Cip1 Antibody
Catalog: DF6423
Description: Rabbit polyclonal antibody to p21 Cip1
Application: WB IHC IF/ICC
Cited expt.: WB, IHC, IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Horse, Rabbit, Dog
Mol.Wt.: 21kDa; 18kD(Calculated).
Uniprot: P38936
RRID: AB_2838386

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user. For optimal experimental results, antibody reuse is not recommended.
*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:
Horse(83%), Rabbit(100%), Dog(83%)
Clonality:
Polyclonal
Specificity:
p21 Cip1 Antibody detects endogenous levels of total p21 Cip1.
RRID:
AB_2838386
Cite Format: Affinity Biosciences Cat# DF6423, RRID:AB_2838386.
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:

A synthesized peptide derived from human p21 Cip1, corresponding to a region within the internal amino acids.

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:
The tumor suppressor protein p21 Waf1/Cip1 acts as an inhibitor of cell cycle progression. It functions in stoichiometric relationships forming heterotrimeric complexes with cyclins and cyclin-dependent kinases. In association with CDK2 complexes, it serves to inhibit kinase activity and block progression through G1/S (1). However, p21 may also enhance assembly and activity in complexes of CDK4 or CDK6 and cyclin D (2). The carboxy-terminal region of p21 is sufficient to bind and inhibit PCNA, a subunit of DNA polymerase, and may coordinate DNA replication with cell cycle progression (3). Upon UV damage or during cell cycle stages when cdc2/cyclin B or CDK2/cyclin A is active, p53 is phosphorylated and upregulates p21 transcription via a p53-responsive element (4). Protein levels of p21 are downregulated through ubiquitination and proteasomal degradation (5).
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
Rabbit
100
Horse
83
Dog
83
Pig
0
Bovine
0
Sheep
0
Xenopus
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

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.

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). The NLRX1-SLC39A7 complex orchestrates mitochondrial dynamics and mitophagy to rejuvenate intervertebral disc by modulating mitochondrial Zn2+ trafficking. Autophagy, 2024 (PubMed: 37876250) [IF=14.6]

Application: WB    Species: human    Sample: NP cells

Figure 1. Downregulation of NLRX1 correlates with aggravated human NP cell senescence and IDD progression. Human NP tissue specimens with different degenerative grades were collected for histological analysis. (A) Representative MRI images at T2 weight sequence were evaluated by Pfirrmann grading system. II: grade II, III: grade III, IV: grade IV. (B) histological analysis of human NP samples by alcian blue staining, scale bar: 100 μm. (C) immunohistochemical staining of CDKN2A, MKI67 and NLRX1 in different degenerative NP tissues, scale bar: 100 μm. (D and E) linear regression analyses of the tissue staining intensity of CDKN2A and that of NLRX1 (D), or the intensity of MKI67 and that of NLRX1 (E). AOD, average optical density. (F-H) protein expressions of senescence indicators (TP53, CDKN1A, CDKN2A), SASP factors (IL1B, IL6) and NLRX1 in primary human NP cells isolated from different degenerative NP tissues with the treatment of TBHP (100 μM), as determined by western blotting. (I-L) cell senescence (SA-GLB1/β-gal staining), cell proliferation (EdU incorporation) and NLRX1 expression (immunofluorescent staining) in primary human NP cells isolated from different degenerative NP tissues with the treatment of TBHP (100 μM), scale bar: 100 μm. (M and N) MRI examination, hematoxylin and eosin (HE) and safranin-O (SO) staining in sham or operation-induced degenerated disc of rat, scale bar: 500 μm (left panel), 50 μm (right panel). (O and P) immunohistochemical staining of aggrecan, collagen type II, NLRX1 and CDKN2A in sham or operation-induced degenerated disc of rat, scale bar: 500 μm (left panel), 50 μm (right panel). Data are represented as mean ± SD.
2). Complement C4b as a key mediator of synaptic loss and cognitive decline in brain aging. Molecular therapy : the journal of the American Society of Gene Therapy, 2025 (PubMed: 40931518) [IF=12.1]
3). Multiomic identification of senescent stem cell populations critical for osteoarthritis progression and therapy in subchondral bones. Science advances, 2025 (PubMed: 40680126) [IF=11.7]

Application: IF/ICC    Species: human    Sample:

Fig. 3. Spatial transcriptome analysis of the I~II and III~IV side tissues from the patients with OA. (A) the OA signature gene score in the articular surface (AS), super-ficial zone (SZ), middle zone (MZ), and deep zone (dZ) groups. (B) the aging score in the AS, SZ, MZ, and dZ groups. (C) dot plot showing the expression of the senescencegenes in the AS, SZ, MZ, and dZ groups. the size of the dots represents the percentage of genes expressed in the AS, SZ, MZ, and dZ groups. the color of the dot representsthe average expression of the gene. pct.exp, percent expressed. (D) violin plot showing the expression of the representative senescence genes in the AS, SZ, MZ, and dZgroups. (E) Box plot showing the aging score between healthy people and patients with OA in the AS, SZ, MZ, and dZ groups. (F) violin plot showing the expression of theEGFR gene in the AS, SZ, MZ, and dZ groups. (G) Box plot showing the expression of the EGFR gene between healthy people and patients with OA in the AS, SZ, MZ, anddZ groups. (H) Multiplex immunofluorescence staining showing the co-expression of p21, p16, and eGFR in the i~ii and iii~iv side subchondral bone tissues from patientswith OA. dAPi, 4′,6-diamidino- 2- phenylindole. (I) Semiquantitative analysis from five randomly selected low-power fields showing the cell proportion of eGFR+p21+ cellsin each group (n = 5 donors). (J) Semiquantitative analysis from five randomly selected low-power fields showing the cell proportion of eGFR+p16+ cells in each group(n = 5 donors). *P < 0.05; **P < 0.01; ***P < 0.001; n.s., not significant. SB, subchondral bone; StB, subchondral trabecular bone. Ucell, a package for evaluating genesignatures based on the Mann-Whitney U statistic.
4). SYNJ2BP ameliorates intervertebral disc degeneration by facilitating mitochondria-associated endoplasmic reticulum membrane formation and mitochondrial Zn2+ homeostasis. Free radical biology & medicine, 2024 (PubMed: 38158052) [IF=7.1]

Application: WB    Species: human    Sample: NP cell

Fig. 1. MAM structure is impaired in degenerated disc. (A) Confocal analysis of TOMM20 and CANX with IF staining in human NP cells isolated from health or degenerated NP tissues, scale bar: 10 μm. (B) MAM structure analysis by TEM in human NP cells isolated from health or degenerated NP tissues, scale bar: 1 μm. (C, D) Protein expressions of cell senescence indicators (TP53, CDKN1A, CDKN2A) and extracellular matrix (COL2A1) in primary human NP cells isolated from health NP tissues with the treatment of TBHP, as determined by western blotting. (E-G) Cell senescence (SA-GLB1/β-gal staining) and cell proliferation (EdU incorporation) in primary human NP cells isolated from health NP tissues with the treatment of TBHP, scale bar: 100 μm (white light images), 200 μm (IF images). (H) Confocal analysis of TOMM20 and CANX with IF staining in human NP cells isolated from health NP tissues with the treatment of TBHP, scale bar: 10 μm. (I) MAM structure analysis by transmission electron microscopy (TEM) in human NP cells isolated from health NP tissues with the treatment of TBHP, scale bar: 1 μm. Data are represented as mean ± SD. *p 
5). Kang Shuai Lao Pian exerts anti-aging effects by enhancing mitochondrial oxidative metabolism. Phytomedicine : international journal of phytotherapy and phytopharmacology, 2025 (PubMed: 41101068) [IF=6.7]
6). PIM3 enhances nucleus pulposus cell function and extracellular matrix integrity via kinase-dependent activation of the Akt/mTOR signaling axis in intervertebral disc degeneration. Translational research : the journal of laboratory and clinical medicine, 2026 (PubMed: 41478528) [IF=6.4]
7). The role of miR-217/SIRT1 Axis in modulating the NLRP3 Inflammasome in the naturally aging thoracic aorta and in endothelial cells undergoing senescence induced by H2O2. International immunopharmacology, 2025 (PubMed: 41027061) [IF=4.8]
8). Specnuezhenide Alleviates Senile Osteoporosis by Activating TGR5/FXR Signaling in Bone Marrow Mesenchymal Stem Cells and RANKL-Induced Osteoclasts. Drug design, development and therapy, 2025 (PubMed: 40066080) [IF=4.7]

Application: IHC    Species: Mouse    Sample:

Figure 4 SPN attenuated senescence in D-gal-induced mice. (A) Immunohistochemistry of P16, P21, P53 and (B) related statistical analysis (n=5). Data are expressed as the mean ± SD. **P < 0.01 compared with the MOD group.
9). Crocetin inhibits the proliferation, migration and TGF-β 2-induced epithelial-mesenchymal transition of retinal pigment epithelial cells . European Journal of Pharmacology, 2017 (PubMed: 28970011) [IF=4.2]

Application: WB    Species: human    Sample:

Figure 2. Crocetin induces cell cycle arrest and influences PCNA, p21, and p53 expression in RPE cells. (A) Cells were harvested after treatment with or without 50 and 100 μM crocetin for 24 h. DNA was stained with PI for flow cytometric analysis. The number of cells in G1 phase was significantly increased in the crocetin-treated group compared with that in the untreated group. (B) Cells were treated or without with 50, 100 and 200 μM crocetin for 48 h. Western blot analysis was used to evaluated the expression of PCNA, p21, p53 and the housekeeping protein β-actin. *P< 0.05 vs 0 μM crocetin, **P< 0.01 vs 0 μM crocetin. The data are presented as the mean ± S.D. (n = 3/group).
10). Hydrogen promoted mitochondrial autophagy and alleviated CIH-induced vascular endothelial cell senescence by regulating oxidative stress. European journal of pharmacology, 2025 (PubMed: 40845957) [IF=4.2]
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