Antibodies, Proteins, Kits and Reagents for Life Science

361 843 ₸ Product size

100 µl 361 843 ₸

Order now and get it on Tuesday March 02, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR3993] to p21
Suitable for: WB
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-p21 antibody [EPR3993]
See all p21 primary antibodies
Description

Rabbit monoclonal [EPR3993] to p21
Host species

Rabbit
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 samples. We recommended ab109520 and ab188224 for use in IHC.
Tested Applications & Species

Application Species
WB
Mouse

Rat

Human

See all applications and species data
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HUVEC, U87-MG, MCF-7, LnCaP, RAW264.7, PC-12, HT1080, HCT116 and C6 cell lysates; Wild-type DLD-1 20 µM 2,3-DCPE for 16hrs treated cell lysate. ICC/IF: HeLa cells.
General notes
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Application	Species
WB	Mouse Rat Human

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR3993
Isotype

IgG
Research areas

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, 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 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 (ab216773) and 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.
Western blot - Anti-p21 antibody [EPR3993] (ab109199)

All lanes : Anti-p21 antibody [EPR3993] (ab109199) at 1/1000 dilution

Lane 1 : Wild-type HCT116 cell lysate
Lane 2 : CDKN1A knockout HCT116 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 18 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab109199 observed at 20 kDa. Red - 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 ab266860 (knockout 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 (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 (ab216773) and 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.
Western blot - Anti-p21 antibody [EPR3993] (ab109199)

Lane 1 : Anti-p21 antibody [EPR3993] (ab109199) (0.7ug/ul)
Lane 2 : Anti-p21 antibody [EPR362] (ab109520) (0.8ug/ul)

All lanes : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 18 kDa

Exposure time: 3 minutes

Blocking and diluting buffer: 5% NDFM/TBST.
Western blot - Anti-p21 antibody [EPR3993] (ab109199)

Lane 1 : Anti-p21 antibody [EPR3993] (ab109199) (1.4ug/ul)
Lane 2 : Anti-p21 antibody [EPR18021] (ab188224) (1.0ug/ul)

Lane 1 : RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
Lane 2 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : 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, ab8245, observed at 37 kDa.

This western blot image is a comparison between ab109119 and a competitor's top cited rabbit polyclonal antibody.
Western blot - Anti-p21 antibody [EPR3993] (ab109199)

Anti-p21 antibody [EPR3993] (ab109199) at 1/1000 dilution (purified) + PC-12 cell lysate at 10 µg

Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 18 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Anti-p21 antibody [EPR3993] (ab109199)

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