WB
Western blot analysis using p21 antibody against Nih3t3(1),C2C12(2) lysate . 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG (H&L) - HRP(OM644366), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60SIHC
Immunohistochemical analysis of paraffin-embedded bladder cancer tissues using p21 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:400 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded cervical cancer tissues using p21 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:400 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded ovarian cancer tissues using p21 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:400 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded rectal cancer tissues using p21 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:400 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.ICC/IF
Immunofluorescence analysis of U251 cells usingp p21 antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 15 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Mouse IgG(H&L) (Green,OM643485), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutesELISA
Blue line: Antibody; Orange line: Isotype control antibodyWB
Western blot analysis using p21 antibody against Hela(1),Cos-7(2) cell . 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG (H&L) - HRP(OM644366), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S.| Product Name | Anti-p21 antibody,pbs only |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Mouse |
| Tested Applications | ELISAICC/IFIHCWB |
| WB:1:1000-1:2000 IHC:1:200-1:2000 ICC/IF:1:100-1:500 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | CDKN1A |
|---|---|
| Gene Synonyms | CAP20 CDKI CDKN1 CIP1 MDA-6 mda6 P21 p21CIP1 p21WAF SDI1 WAF1 |
| Gene Full Name | cyclin dependent kinase inhibitor 1A |
| Gene Summary | This gene encodes a potent cyclin-dependent kinase inhibitor. The encoded protein binds to and inhibits the activity of cyclin-cyclin-dependent kinase2 or -cyclin-dependent kinase4 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. This protein can interact with proliferating cell nuclear antigen, a DNA polymerase accessory factor, and plays a regulatory role in S phase DNA replication and DNA damage repair. This protein was reported to be specifically cleaved by CASP3-like caspases, which thus leads to a dramatic activation of cyclin-dependent kinase2, and may be instrumental in the execution of apoptosis following caspase activation. Mice that lack this gene have the ability to regenerate damaged or missing tissue. Multiple alternatively spliced variants have been found for this gene. [provided by RefSeq, Sep 2015] |
| Alternative Names | Activating Fragment 1 alternate gene name=WAF1 CDK-interacting protein 1 CDK-interaction protein 1 CDKN1A CDNK1A Cip1) cyclin kinase inhibitor Cyclin-dependent kinase inhibitor 1A (p21 cyclin-dependent kinase inhibitor 1A (p21 Cip1) DNA synthesis inhibitor melanoma differentiation associated protein 6 melanoma differentiation-associated protein OTTHUMP00000016298 p21Cip1/Waf1 putative DNA synthesis inhibitor SLC12A9 Wild type p53 activated fragment 1 (WAF1) wild type p53 activated fragment-1 wild-type p53-activated fragment 1 |
| Molecular Weight(MW) | 21kDa(observed:35、21kDa) |
| Cellular Localization | Cytoplasm,Nucleus. |
| SwissProt ID | P38936 |
|---|
WB
Western blot analysis using p21 antibody against Nih3t3(1),C2C12(2) lysate . 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG (H&L) - HRP(OM644366), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S
IHC
Immunohistochemical analysis of paraffin-embedded bladder cancer tissues using p21 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:400 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded cervical cancer tissues using p21 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:400 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded ovarian cancer tissues using p21 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:400 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded rectal cancer tissues using p21 antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:400 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG(H&L)-HRP (OM644366), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
ICC/IF
Immunofluorescence analysis of U251 cells usingp p21 antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 15 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Mouse IgG(H&L) (Green,OM643485), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutes
ELISA
Blue line: Antibody; Orange line: Isotype control antibody
WB
Western blot analysis using p21 antibody against Hela(1),Cos-7(2) cell . 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Mouse IgG (H&L) - HRP(OM644366), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S.| Application Notes | WB:1:1000-1:2000 IHC:1:200-1:2000 ICC/IF:1:100-1:500 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. |
| Storage Buffer | PBS ONLY |