Anti-P21/CDKN1A Antibody Picoband™

Product Name

Anti-P21/CDKN1A Antibody Picoband™

View all p21/CIP1/CDKN1A Antibodies

SKU/Catalog Number

A00145-1

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-P21/CDKN1A Antibody Picoband™ catalog # A00145-1. Tested in ELISA, IF, IHC, ICC, WB applications. This antibody reacts with Human.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-P21/CDKN1A Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00145-1)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human P21/CDKN1A recombinant protein (Position: M1-R140).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A00145-1 is reactive to CDKN1A in Human

Applications

A00145-1 is guaranteed for ELISA, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

21 kDa

Calculated molecular weight

18.119kDa

Background of p21/CIP1/CDKN1A

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.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 µg/ml, Human
Immunohistochemistry(Paraffin-embedded Section), 2-5 µg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Direct ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of P21/CDKN1A using anti-P21/CDKN1A antibody (A00145-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: human U20S whole cell lysates,
Lane 5: human SIHA whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-P21/CDKN1A antigen affinity purified polyclonal antibody (Catalog # A00145-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for P21/CDKN1A at approximately 21 kDa. The expected band size for P21/CDKN1A is at 21 kDa.

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Figure 2. IHC analysis of P21/CDKN1A using anti-P21/CDKN1A antibody (A00145-1).
P21/CDKN1A was detected in a paraffin-embedded section of human thyroid cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-P21/CDKN1A Antibody (A00145-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 3. IHC analysis of P21/CDKN1A using anti-P21/CDKN1A antibody (A00145-1).
P21/CDKN1A was detected in a paraffin-embedded section of human ovarian cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-P21/CDKN1A Antibody (A00145-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 4. IF analysis of P21/CDKN1A using anti-P21/CDKN1A antibody (A00145-1) and anti-Beta Tubulin antibody (M01857-3).
P21/CDKN1A was detected in immunocytochemical section of Hela cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-P21/CDKN1A Antibody (A00145-1) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight®488 Goat Anti-Rabbit IgG (BA1127) and DyLight®550 Conjugated Goat Anti-Mouse IgG (BA1133) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Gene/Protein Information For CDKN1A (Source: Uniprot.org, NCBI)

Alternative Names

CAP20cyclin-dependent kinase inhibitor 1; CDK-interacting protein 1; CDKN1A; CDKN1melanoma differentiation associated protein 6; CIP1; CIP1WAF1CDK-interaction protein 1; cyclin-dependent kinase inhibitor 1A (p21, Cip1); MDA6; MDA-6; Melanoma differentiation-associated protein 6; p21; p21CIP1; p21Cip1/Waf1; PIC1; SDI1DNA synthesis inhibitor; wild-type p53-activated fragment 1 CDKN1A CAP20, CDKN1, CIP1, MDA-6, P21, SDI1, WAF1, p21CIP1 cyclin dependent kinase inhibitor 1A cyclin-dependent kinase inhibitor 1|CDK-interacting protein 1|CDK-interaction protein 1|DNA synthesis inhibitor|cyclin-dependent kinase inhibitor 1A (p21, Cip1)|melanoma differentiation associated protein 6|wild-type p53-activated fragment 1

For more info on CDKN1A, check out the CDKN1A Infographic

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In this infographic, you will see the following information for CDKN1A: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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