Name: Anti-p21 antibody [EPR3993]
SKU: ab109199
Rating: 4 (12 reviews)

Anti-p21 antibody [EPR3993] (ab109199) is a rabbit monoclonal antibody detecting p21 in Western Blot. Suitable for Human, Mouse, Rat.

- KO validated for confirmed specificity

- Biophysical QC for unrivalled batch-batch consistency

- Over 300 publications

Images

Western blot - Anti-p21 antibody [EPR3993] (AB109199), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB
Human	Tested
Mouse	Tested
Rat	Tested

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes For unpurified use at 1/1000 - 1/10000. Not recommended for use with tissue samples.
Species Rat	Dilution info 1/1000	Notes For unpurified use at 1/1000 - 1/10000. Not recommended for use with tissue samples.
Species Human	Dilution info 1/1000	Notes For unpurified use at 1/1000 - 1/10000. Not recommended for use with tissue samples.

Associated Products

Select an associated product type

9 products for Alternative Product

Target data

See full target information CDKN1A

Function

Plays an important role in controlling cell cycle progression and DNA damage-induced G2 arrest (PubMed:9106657). Involved in p53/TP53 mediated inhibition of cellular proliferation in response to DNA damage. Also involved in p53-independent DNA damage-induced G2 arrest mediated by CREB3L1 in astrocytes and osteoblasts (By similarity). 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). Negatively regulates the CDK4- and CDK6-driven phosphorylation of RB1 in keratinocytes, thereby resulting in the release of E2F1 and subsequent transcription of E2F1-driven G1/S phase promoting genes (By similarity).

Alternative names

CAP20, CDKN1, CIP1, MDA6, PIC1, SDI1, WAF1, CDKN1A, Cyclin-dependent kinase inhibitor 1, CDK-interacting protein 1, Melanoma differentiation-associated protein 6, p21, MDA-6

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR3993
Purification technique: Affinity purification Protein A
Specificity: Expression levels of the target protein vary between different tissue/cell lines and in some cases, induction may be required before a signal is observed.
This antibody is not recommended for use in WB with tissue and primary cell samples.
We recommended ab109520 and ab188224 for use in IHC.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Stable for 12 months at -20°C

Notes

Product Specifications
Anti-p21 antibody [EPR3993] (ab109199) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in WB in human, mouse, rat samples.
Anti-p21 antibody [EPR3993] (ab109199) specifically detects p21 (UniProt ID: P38936; Molecular weight: 18kDa) and is sold in 100 µL and 1 mL selling sizes.

Quality and Validation
Abcam's high quality manufacturing and validation processes ensure Anti-p21 antibody [EPR3993] (ab109199) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
The specificity of Anti-p21 antibody [EPR3993] (ab109199) has been confirmed by testing in knockout samples.
Anti-p21 antibody [EPR3993] (ab109199) has been cited over 305 times in peer reviewed journals and is trusted by the scientific community.
Anti-p21 antibody [EPR3993] (ab109199) has 10 independent reviews from customers.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

10 product images

Western blot - Anti-p21 antibody [EPR3993] (ab109199)
False colour image of Western blot: Anti-p21 antibody [EPR3993] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109199 was shown to bind specifically to p21. A band was observed at 21 kDa in wild-type HeLa cell lysates with no signal observed at this size in CDKN2A knockout cell line Human CDKN1A knockout HeLa cell line ab255349 (knockout cell lysate ab263812). To generate this image, wild-type and CDKN2A knockout cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-p21 antibody [EPR3993] (ab109199) at 1/1000 dilution
Lane 1: wild-type HeLa Vehicle Control Fluvastatin (0 uM, 24 h) cell lysate at 20 µg
Lane 2: wild-type HeLa Treated Fluvastatin (50 uM, 24 h) cell lysate at 20 µg
Lane 2: Western blot - Human CDKN1A knockout HeLa cell line (Human CDKN1A knockout HeLa cell line ab255349)
Lane 3: CDKN1A knockout HeLa Vehicle Control Fluvastatin (0 uM, 24 h) cell lysate at 20 µg
Lane 4: CDKN1A knockout HeLa Treated Fluvastatin (50 uM, 24 h) cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 18 kDa
Observed band size: 21 kDa
Western blot - Anti-p21 antibody [EPR3993] (ab109199)
Lane 1: Wild-type DLD-1 cell lysate (20 μg)
Lane 2: Wild-type DLD-1 20 μM 2,3-DCPE for 16hrs treated cell lysate (20 μg)
Lane 3: p21 knockout DLD-1 cell lysate (20 μg)
Lane 4: p21 knockout 20 μM 2,3-DCPE for 16hrs DLD-1 cell lysate (20 μg)
Lane 5: HT1080 cell lysate (20 μg)

Lanes 1 - 5: Merged signal (red and green). Green - ab109199 observed at 20 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
ab109199 was shown to recognize p21 in WT DLD-1 cells with 2,3-DCPE treatment along with additional cross-reactive bands. When p21 knockout DLD-1 cells +/- 2,3-DCPE treatment were used, no band was observed. Wild-type and p21 knockout samples were subjected to SDS-PAGE. ab109199 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-p21 antibody [EPR3993] (ab109199)
Predicted band size: 18 kDa
Observed band size: 20 kDa
Western blot - Anti-p21 antibody [EPR3993] (ab109199)
Lanes 1- 2: Merged signal (red and green). Green - ab109199 observed at 20 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
ab109199 was shown to react with p21 in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line Human CDKN1A knockout HCT116 cell line ab266860 (knockout cell lysate Human CDKN1A knockout HCT116 cell lysate ab256870) was used. Wild-Type HCT116 and CDKN1A knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109199 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-p21 antibody [EPR3993] (ab109199) at 1/1000 dilution
Lane 1: Wild-type HCT116 cell lysate at 20 µg
Lane 2: CDKN1A knockout HCT116 cell lysate at 20 µg
Lane 2: Western blot - Human CDKN1A knockout HCT116 cell line (Human CDKN1A knockout HCT116 cell line ab266860)
Performed under reducing conditions.
Predicted band size: 18 kDa, 197 kDa, 36 kDa, 47 kDa, 72 kDa, 84 kDa
Observed band size: 150 kDa, 20 kDa, 40 kDa, 45 kDa, 47 kDa
Western blot - Anti-p21 antibody [EPR3993] (ab109199)
Different batches of ab109199 were tested on MCF7 (Human breast adenocarcinoma epithelial cell) lysate at 0.2 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 21 kDa.
All lanes: Western blot - Anti-p21 antibody [EPR3993] (ab109199)
Predicted band size: 18 kDa
Western blot - Anti-p21 antibody [EPR3993] (ab109199)
Blocking and diluting buffer: 5% NDFM/TBST.
All lanes: Western blot - Anti-p21 antibody [EPR3993] (ab109199)
All lanes: MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 18 kDa
Exposure time: 3min
Western blot - Anti-p21 antibody [EPR3993] (ab109199)
All lanes: Western blot - Anti-p21 antibody [EPR3993] (ab109199)
Lane 1: RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates at 15 µg
Lane 2: Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 18 kDa
Western blot - Anti-p21 antibody [EPR3993] (ab109199)
Lane 1: Wild-type DLD-1 cell lysate (20 μg)
Lane 2: Wild-type DLD-1 20 μM 2,3-DCPE for 16hrs treated cell lysate (20 μg)
Lane 3: p21 knockout DLD-1 cell lysate (20 μg)
Lane 4: p21 knockout 20 μM 2,3-DCPE for 16hrs DLD-1 cell lysate (20 μg)
Lane 5: HT1080 cell lysate (20 μg)

Lanes 1 - 5: Merged signal (red and green). Green - ab109199 observed at 20 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
This western blot image is a comparison between Anti-PIST antibody [EPR4079] ab109119 and a competitor's top cited rabbit polyclonal antibody.
All lanes: Western blot - Anti-p21 antibody [EPR3993] (ab109199)
Predicted band size: 18 kDa
Western blot - Anti-p21 antibody [EPR3993] (ab109199)
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-p21 antibody [EPR3993] (ab109199) at 1/1000 dilution
All lanes: PC-12 cell lysate at 10 µg
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 18 kDa
Observed band size: 21 kDa
Western blot - Anti-p21 antibody [EPR3993] (ab109199)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as a GAPDH loading control.

We recommend using higher or super higher sensitivity ECL substrate for detecting.

Increase lysate amount can also help to get stronger signal.
All lanes: Western blot - Anti-p21 antibody [EPR3993] (ab109199) at 1/1000 dilution
Lane 1: PC-12(Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
Lane 2: PC-12(Rat adrenal gland pheochromocytoma) treated with 50ng/ml NFG for 48 hours whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 18 kDa
Observed band size: 18 kDa
Exposure time: 180s
Western blot - Anti-p21 antibody [EPR3993] (ab109199)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as a GAPDH loading control.
All lanes: Western blot - Anti-p21 antibody [EPR3993] (ab109199) at 1/1000 dilution
Lane 1: Raw 264.7(Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 2: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 3: C6 (Rat glial tumor glial cell) whole cell lysate at 20 µg
Lane 4: PC-12(Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 18 kDa
Observed band size: 18 kDa
Exposure time: 180s