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  • Product Name
    Phospho-p21 Cip1 (Thr145) Antibody
  • Catalog No.
    AF3290
  • RRID
    AB_2834711
  • Source
    Rabbit
  • Application
    WB,IHC,IF/ICC,ELISA
  • Reactivity
    Human, Mouse, Rat
  • Prediction
    Bovine(100%), Horse(91%), Sheep(100%), Rabbit(83%), Dog(100%)
  • UniProt
  • Mol.Wt
    (Observed)21kD,25kD; (Calculated)18kDa
  • Concentration
    1mg/ml
  • Browse similar products>>

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Product Information

Alternative Names:Expand▼

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;

Applications:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000

Reactivity:

Human, Mouse, Rat

Predicted Reactivity:

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

Source:

Rabbit

Clonality:

Polyclonal

Purification:

The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.

Specificity:

Phospho-p21 Cip1 (Thr145) Antibody detects endogenous levels of p21 Cip1 only when phosphorylated at Threonine 145.

RRID:

AB_2834711
Please cite this product as: Affinity Biosciences Cat# AF3290, RRID:AB_2834711.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

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.

Immunogen Information

Immunogen:

A synthesized peptide derived from human p21 Cip1 around the phosphorylation site of Thr145.

Uniprot:



>>Visit The Human Protein Atlas

Gene ID:

Gene Name:

CDKN1A

Molecular Weight:

Observed Mol.Wt.: (Observed)21kD,25kD.
Predicted Mol.Wt.: (Calculated)18kDa.

Subcellular Location:

Cytoplasm. Nucleus.

Tissue Specificity:

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

Research Background

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 (PubMed:11595739). Plays an important role in controlling cell cycle progression and DNA damage-induced G2 arrest (PubMed:9106657).

Post-translational Modifications:

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 assertionGraphics by Christian Stolte

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 (PubMed:19536131). Interacts with PSMA3 (PubMed:11350925). Interacts with PCNA (PubMed:11595739, PubMed:18794347, PubMed:18703516, PubMed:8861913). 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 (PubMed:9106657). 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 (PubMed:19445729). Interacts with PIM1 (PubMed:12431783). Interacts with STK11 and NUAK1 (PubMed:25329316). Interacts wih DTL (PubMed:23213251). Interacts with isoform 1 and isoform 2 of TRIM39 (PubMed:23213251).

Similarity:

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

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 > FoxO signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Jak-STAT signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.(View pathway)
· Human Diseases > Cancers: Specific types > Pancreatic cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Basal cell carcinoma.(View pathway)
· Human Diseases > Cancers: Specific types > Renal cell carcinoma.(View pathway)
· Human Diseases > Cancers: Specific types > Thyroid cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Bladder cancer.(View pathway)
· Human Diseases > Cancers: Overview > Pathways in cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Gastric cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Non-small cell lung cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Glioma.(View pathway)
· Human Diseases > Cancers: Specific types > Colorectal cancer.(View pathway)
· Human Diseases > Cancers: Overview > Proteoglycans in cancer.
· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma.(View pathway)
· Human Diseases > Cancers: Overview > MicroRNAs in cancer.
· Human Diseases > Cancers: Specific types > Endometrial cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Breast 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 > Infectious diseases: Viral > Hepatitis C.
· Human Diseases > Cancers: Overview > Viral carcinogenesis.
· Human Diseases > Infectious diseases: Viral > Hepatitis B.
· Human Diseases > Cancers: Specific types > Melanoma.(View pathway)
· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.
· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.
· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.
· Human Diseases > Infectious diseases: Viral > HTLV-I infection.
· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.
· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.
· Human Diseases > Cancers: Specific types > Prostate cancer.(View pathway)
· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

Reference Citations:

1). Wang Y et al. Cholesterol regulates cell proliferation and apoptosis of colorectal cancer by modulating miR-33a-PIM3 pathway. Biochem Biophys Res Commun 2019 Apr 9;511(3):685-692 (PubMed: 30827510)

2). Yang Q et al. Methyl-β-cyclodextrin potentiates the BITC-induced anti-cancer effect through modulation of the Akt phosphorylation in human colorectal cancer cells. Biosci Biotechnol Biochem 2018 Dec;82(12):2158-2167 (PubMed: 30200817)

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Catalog Number :

AF3290-BP
(Blocking peptide available as AF3290-BP)

Price/Size :

$350/1mg.
Tips: For phospho antibody, we provide phospho peptide(0.5mg) and non-phospho peptide(0.5mg).

Function :

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. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Sheep
100%
Bovine
100%
Dog
100%
Horse
91%
Rabbit
83%
Zebrafish
67%
Xenopus
56%
Pig
0%
Chicken
0%
High similarity Medium similarity Low similarity No similarity
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
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.

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