Invitrogen

Phospho-p21 (Thr145) Polyclonal Antibody

Product Image Gallery

Phospho-p21 (Thr145) Antibody — Figure 5 PRMT6-mediated p21 methylation affects p21 phosphorylation and chemosensitivity of cancer cells A. Wild-type or arginine 156-substituted p21 and PRMT6 were co-overexpressed in 293T cells and phosphorylation levels of the p21 proteins at threonine 145 were examined following immunoprecipitation. The signal intensity of phosphor-p21 (Thr 145) was quantified and normalized by p21 amount. B. Threonine 145-substituted p21 mutant (p21-T145A) or threonine 145/arginine 156 double substituted p21 mutant (p21-T145A/R156A) and PRMT6 were co-overexpressed in HCT 116 p53 +/+ cells. Nuclear and cytoplasmic fractions were prepared and analyzed by western blot. The signal intensity of p21 bands in each fraction was quantified and normalized by that of alpha-Tubulin or histone H3. C. HCT 116 p53 +/+ (left) or HCT116 p53 -/- (right) cells were transfected with mock or PRMT6 expression vector and incubated for 40 h. Subsequently, the cells were further incubated in the presence of 0.5 muM doxorubicin for additional 8 h. Cell cycle distribution was analyzed by flow cytometry coupled with BrdU staining. P -values were calculated with Student and apos;s t test (** P < 0.01, * P < 0.05). D. HCT116 p53 +/+ cells transfected with a mock vector or a PRMT6 expression vector were treated with various concentrations of doxorubicin 24 h post transfection. Cell viability was measured 96 h after the drug treatment. IC50 was calculated using the SigmaPlot software.

Phospho-p21 (Thr145) Antibody in Immunohistochemistry (IHC) — Immunohistochemical analysis of formalin-fixed, paraffin-embedded human cancer tissue using a Phospho-P21CIP1 pThr145 polyclonal antibody (Product # PA5-12646), followed by HRP-conjugated secondary antibody and AEC staining.

Phospho-p21 (Thr145) Antibody in Western Blot (WB) — Western blot analysis of Phospho-P21CIP1 pThr145 using a Phospho-P21CIP1 pThr145 polyclonal antibody (Product # PA5-12646) in HeLa tissue lysate.

Product Details

Tested Applications

Dilution

Dot blot (DB)

1:500

Immunohistochemistry (Paraffin) (IHC (P))

1:50-1:100

Western Blot (WB)

1:1000

Published Applications

Miscellaneous PubMed (MISC)

See 1 publication below

Western Blot (WB)

See 1 publication below

Product Specifications

Species Reactivity

Human

Published species

Human

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

KLH conjugated synthetic phosphopeptide corresponding to amino acid residues surrounding T145 of human P21CIP1

Conjugate

Unconjugated

Form

Liquid

Concentration

0.45 mg/mL

Purification

Protein A, Antigen affinity chromatography

Storage buffer

PBS

Contains

0.09% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_10979470

Target Information

Protein p21 is a tumor suppressor protein which is an inhibitor of Cyclin-dependent kinases (CDK) and is transcriptionally activated by p53. P21 is important in the response of cells to genotoxic stress and a major transcriptional target of p53 protein. The occurrence of p21 in the nucleus executes binding and inhibition activity of cyclin dependent kinases Cdk1 and Cdk2, and blocks the transition from G1 phase into S phase or from G2 phase into mitosis after DNA damage, which enables the repair of damaged DNA. In the cytoplasm, p21 protein has an anti-apoptotic effect. P21 is able to bind to and inhibit caspase 3, as well as the apoptotic kinases ASK1 and JNK. P21 exhibits a dual function in carcinogenesis, and acts as a tumor suppressor, prevents apoptosis, and acts as an oncogene.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: CAP20; CDK-interacting protein 1; CDK-interaction protein 1; CDKN1; CDNK1A; Cyclin-dependent kinase inhibitor 1; cyclin-dependent kinase inhibitor 1A (p21, Cip1); DNA synthesis inhibitor; MDA-6; melanoma differentiation associated protein 6; Melanoma differentiation-associated protein 6; p21; p21CIP1; SDI1; WAF1; wild-type p53-activated fragment 1

Gene Aliases: CAP20; CDKN1; CDKN1A; CIP1; MDA-6; MDA6; P21; p21CIP1; PIC1; SDI1; WAF1

UniProt ID: (Human) P38936

Entrez Gene ID: (Human) 1026

Molecular Function: enzyme modulator kinase inhibitor kinase modulator