p21 Waf1 / Cip1 Antibody (R.229.6)

p21 Waf1 / Cip1 Monoclonal Antibody for Western blot, IF, ICC, IHC (P), FACS, IP

>> See 21 other antibodies for CDKN1A (CDK-interacting protein 1; CDK-interaction protein 1; DNA synthesis inhibitor; OTTHUMP00000016298; cyclin-dependent kinase inhibitor 1; melanoma differentiation associated protein 6; melanoma differentiation-associated protein 6; p21Cip1/Waf1; wild-type p53-activated fragment 1)
Synonyms:
CDK-interacting protein 1; CDK-interaction protein 1; DNA synthesis inhibitor; OTTHUMP00000016298; cyclin-dependent kinase inhibitor 1; melanoma differentiation associated protein 6; melanoma differentiation-associated protein 6; p21Cip1/Waf1; wild-type p53-activated fragment 1
Entrez Gene ID:
UniProt ID:
Details
Host / Isotype: Rabbit / IgG
Class: Monoclonal
Type: Antibody
Clone: R.229.6
Tested Species Reactivity: Human (Hu), Non-human primate (Nhp)
Immunogen: Synthetic peptide corresponding to residues near the carboxy-terminus of human p21.
Ordering Information
Pierce p21 Waf1 / Cip1 Antibody (R.229.6)
Product #
MA5-14949
Size
100 ul
Price
$250.00
Purchase
Add p21 Waf1 / Cip1 Antibody (R.229.6) to your cart.
Tested Applications Dilution *
Western Blot (WB) 1:1000
Immunofluorescence (IF) 1:400
Immunocytochemistry (ICC) 1:400
Immunohistochemistry (Paraffin) (IHC (P)) 1:50
Flow Cytometry (FACS) 1:200
Immunoprecipitation (IP) 1:100
* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Form Information
Form: Liquid
Purification: Affinity chromatography
Storage Buffer: 0.01M HEPES, pH 7.5, with 100µg/ml BSA, 50% glycerol, 0.15M NaCl
Preservative: <0.02% sodium azide
Storage Conditions: -20°C
Product Specific Information
MA5-14949 targets p21 Waf1/Cip1 in FACS, IF, ICC, IHC (P), IP, and WB applications and shows reactivity with Human, and Non-human primate samples.

The MA5-14949 immunogen is synthetic peptide corresponding to residues near the carboxy-terminus of human p21.

MA5-14949 detects p21 Waf1/Cip1 which has a predicted molecular weight of approximately 21 kDa.

Antibodies to this protein (and modification) were previously sold as part of a Thermo Scientific Cellomics High Content Screening Kit. This replacement antibody is now recommended for researchers who need an antibody for high content cell based assays. It has been thoroughly tested and validated for cellular immunofluorescence (IF) applications. Further optimization including the selection of the most appropriate fluorescent Dylight conjugated secondary antibody may have to be performed for your high content assay.
General Information
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. This protein can interact with proliferating cell nuclear antigen (PCNA), 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 CDK2, and may be instrumental in the execution of apoptosis following caspase activation. Two alternatively spliced variants, which encode an identical protein, have been reported.
Product Images
  • p21 Waf1 / Cip1 Antibody (MA5-14949) in WB
    MA5-14949_WesternBlot.jpg

    Western Blot with anti-p21 Waf1 / Cip1 Monoclonal Antibody [R.229.6] (MA5-14949) (MA5-14949_WesternBlot.jpg)

    Western Blot with anti-p21 Waf1 / Cip1 Monoclonal Antibody [R.229.6] (MA5-14949)

    Western blot analysis of p21 Waf1/Cip1 was performed by loading HUVEC and HeLa cell lysates on a SDS-PAGE gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% nonfat dry milk in TBST for 1 hour at room temperature. The membrane was probed with a p21 Waf1/Cip1 monoclonal antibody (Product # MA5-14949) at a dilution of 1:1000 overnight at 4C, followed by an anti-rabbit IgG-HRP secondary antibody. Detection was performed using a chemiluminescent substrate.
  • p21 Waf1 / Cip1 Antibody (MA5-14949) in IF
    p21Waf1-Cip1_MA5-14949_Antibody_Immunofluorescence(IF)_1A.jpg

    Immunofluorescence with anti-p21 Waf1 / Cip1 Monoclonal Antibody [R.229.6] (MA5-14949) (p21Waf1-Cip1_MA5-14949_Antibody_Immunofluorescence(IF)_1A.jpg)

    Immunofluorescence with anti-p21 Waf1 / Cip1 Monoclonal Antibody [R.229.6] (MA5-14949)

    Immunofluorescent analysis of p21 Waf1/Cip1 (red) in MCF7 cells. Formaldehyde fixed cells were blocked with 5% normal goat serum + 0.3% Triton X-100 in PBS for 1 hour at room temperature. Cells were probed with a p21 Waf1/Cip1 monoclonal antibody (Product # MA5-14949) at a dilution of 1:400 overnight at 4C, followed by a fluorescently-conjugated anti-rabbit IgG secondary antibody. Nuceli were counterstained (blue) and cells were visualized by confocal microscopy.
  • p21 Waf1 / Cip1 Antibody (MA5-14949) in IF
    p21_Antibody_MA5-14949_Immunofluorescence_1.jpg

    Immunofluorescence with anti-p21 Waf1 / Cip1 Monoclonal Antibody [R.229.6] (MA5-14949) (p21_Antibody_MA5-14949_Immunofluorescence_1.jpg)

    Immunofluorescence with anti-p21 Waf1 / Cip1 Monoclonal Antibody [R.229.6] (MA5-14949)

    Immunofluorescent analysis of p21 Waf1/Cip1 (green) in HeLa cells either left untreated (left panel) or treated with 5uM Camptothecin for 3 hours (right panel). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a p21 Waf1/Cip1 monoclonal antibody (Product # MA5-14949) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or ToxInsight Instrument at 20X magnification.
  • p21 Waf1 / Cip1 Antibody (MA5-14949) in IHC
    MA5-14949_Immunohistochemistry.jpg

    Immunohistochemistry with anti-p21 Waf1 / Cip1 Monoclonal Antibody [R.229.6] (MA5-14949) (MA5-14949_Immunohistochemistry.jpg)

    Immunohistochemistry with anti-p21 Waf1 / Cip1 Monoclonal Antibody [R.229.6] (MA5-14949)

    Immunohistochemistry was performed on paraffin-embedded HeLa cells. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer for 10 minutes at 95C. Following antigen-retrieval, tissues were incubated in 3% hydrogen peroxide for 10 minutes, washed, and then blocked in 5% normal goat serum in TBST for 1 hour at room temperature. Tissues were stained with a p21 Waf1/Cip1 monoclonal antibody (Product # MA5-14949) at a dilution of 1:50 overnight at 4C. Tissues were washed extensively with TBS + 0.1% Tween-20 and detection was performed using a HRP-conjugated secondary reagent followed by DAB substrate.
  • p21 Waf1 / Cip1 Antibody (MA5-14949) in FACS
    MA5-14949_FlowCytometry.jpg

    Flow Cytometry with anti-p21 Waf1 / Cip1 Monoclonal Antibody [R.229.6] (MA5-14949) (MA5-14949_FlowCytometry.jpg)

    Flow Cytometry with anti-p21 Waf1 / Cip1 Monoclonal Antibody [R.229.6] (MA5-14949)

    Flow cytometric analysis of p21 Waf1/Cip1 on HeLa (red histogram) and MCF7 cells (blue histogram). Cells were harvested, fixed with formaldehyde, permeabilized with methanol, and incubated with a p21 Waf1/Cip1 monoclonal antibody (Product # MA5-14949) at a dilution of 1:200 for 1 hour at room temperature. Cells were stained with a fluorescently-conjugated anti-rabbit IgG secondary antibody and analyzed on a flow cytometer.
  • p21 Waf1 / Cip1 Antibody (MA5-14949) in IP
    MA5-14949_WesternBlot_2.jpg

    Immunoprecipitation with anti-p21 Waf1 / Cip1 Monoclonal Antibody [R.229.6] (MA5-14949) (MA5-14949_WesternBlot_2.jpg)

    Immunoprecipitation with anti-p21 Waf1 / Cip1 Monoclonal Antibody [R.229.6] (MA5-14949)

    Immunoprecipitation of p21 Waf1/Cip1 was performed on HUVEC cells. Antigen-antibody complexes were captured by incubating 200ul of whole cell lysate with 20ul of an immobilized p21 Waf1/Cip1 monoclonal antibody (Product # MA5-14949) overnight on a rocking platform at 4C. The immune complexes were washed extensively and eluted with SDS sample buffer. Samples were resolved on a SDS-PAGE gel, transferred to a membrane, and blocked with 5% nonfat dry milk in TBST for 1 hour at room temperature. The membrane was probed with a p21 Waf1/Cip1 monoclonal antibody (Product # MA5-14949) at a dilution of 1:1000, followed by an anti-rabbit IgG-HRP secondary antibody. Detection was performed using a chemiluminescent substrate.
(This product is for In Vitro experimental use only. Not for resale without express authorization.)
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