Antibodies, Proteins, Kits and Reagents for Life Science

348 441 ₸ Product size

100 µl 348 441 ₸

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 [EPR17343] to p53
Suitable for: WB, ICC/IF
Knockout validated
Reacts with: Human

Product name

Anti-p53 antibody [EPR17343]
See all p53 primary antibodies
Description

Rabbit monoclonal [EPR17343] to p53
Host species

Rabbit
Specificity

Based on the results of the knockout validation testing this antibody may not be suitable for IHC-P, Flow Cyt, IP or ChIP. Please contact our Scientific Support team for additional information.
Tested Applications & Species

Application Species
ICC/IF
Human

WB
Human

See all applications and species data
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: A431, HEK-293, and T-47D whole cell lysates. ICC/IF: Wild-type HAP1 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.

Application	Species
ICC/IF	Human
WB	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 long term. Avoid freeze / thaw cycle.
Storage buffer

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

Protein A purified
Clonality

Monoclonal
Clone number

EPR17343
Isotype

IgG
Research areas

Western blot - Anti-p53 antibody [EPR17343] (ab179477)

Lane 1: Wild-type HAP1 cell lysate (20 µg)

Lane 2: p53 knockout HAP1 cell lysate (20 µg)

Lane 3: A431 cell lysate (20 µg)

Lane 4: Saos-2 cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab179477 observed at 53 kDa. Red - loading control, ab8226, observed at 42 kDa.

ab179477 was shown to specifically react with p53 in wild type HAP1 cells along with additional cross reactive bands. No band was observed with p53 knockout samples were used.

Wild-type and p53 knockout samples were subjected to SDS-PAGE. ab179477 and ab8226 (loading control to beta Actin) were diluted to 1/2000 and 1/1000 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.
Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [EPR17343] (ab179477)

ab179477 staining p53 in wild-type HAP1 cells (top panel) and p53 knockout HAP1 cells (bottom panel). The cells were fixed with methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab179477 at 1/500 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Western blot - Anti-p53 antibody [EPR17343] (ab179477)

All lanes : Anti-p53 antibody [EPR17343] (ab179477) at 1/2000 dilution

Lane 1 : Untreated HCT 116 (Human colorectal carcinoma) whole cell lysate
Lane 2 : HCT 116 (Human colorectal carcinoma) whole cell lysate with 0.5µM Doxorubicin for 24hours whole cell lysate

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

Predicted band size: 44 kDa
Observed band size: 53 kDa
why is the actual band size different from the predicted?
Western blot - Anti-p53 antibody [EPR17343] (ab179477)

Anti-p53 antibody [EPR17343] (ab179477) at 1/20000 dilution + T-47D (Human ductal breast epithelial carcinoma cell line) whole cell lysates at 20 µg

Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 44 kDa
Observed band size: 44,53 kDa why is the actual band size different from the predicted?

Blocking/dilution buffer: 5% NFDM/TBST.
Western blot - Anti-p53 antibody [EPR17343] (ab179477)

All lanes : Anti-p53 antibody [EPR17343] (ab179477) at 1/2000 dilution

Lane 1 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysates
Lane 2 : A431 (Human epidermoid carcinoma) whole cell lysates
Lane 3 : Saos-2 (Human osteosarcoma cell line) whole cell lysates
Lane 4 : HL-60 (Human promyelocytic leukemia cells) whole cell lysates
Lane 5 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 44 kDa
Observed band size: 53, 44 kDa why is the actual band size different from the predicted?

Blocking and Diluting buffer and concentration: 5% NFDM/TBST.
Saos-2, PC-3 and HL-60 cells are p53 null cell lines.
Anti-p53 antibody [EPR17343] (ab179477)

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