Antibodies, Proteins, Kits and Reagents for Life Science

335 040 ₸ Product size

100 µl 335 040 ₸ 10 µl 137 366 ₸

Order now and get it on Tuesday March 09, 2021

Anti-Mutant p53 antibody [E47] (ab32509)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [E47] to Mutant p53
Suitable for: IHC-P, ICC/IF, WB, IP, Flow Cyt
Reacts with: Human

Product name

Anti-Mutant p53 antibody [E47]
See all Mutant p53 primary antibodies
Description

Rabbit monoclonal [E47] to Mutant p53
Host species

Rabbit
Specificity

ab32509 recognises human mutant forms of p53 but not human p53 wild type. ab32059 showed a negative signal on wildtype p53 cell lines (HepG2, A549, MCF-7) and a positive signal on mutant p53 cell lines (T47-D, Raji, A431) in WB.

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide within Human Mutant p53 aa 350 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P04637
Positive control
- WB: A431 and T-47D cell lysate ICC/IF: A431 cells. IHC-P: Human lung carcinoma tissue. Flow Cyt: HEK-293 cells IP: HEK-293 cell lysates
General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Clonality

Monoclonal
Clone number

E47
Isotype

IgG
Research areas

Western blot - Anti-Mutant p53 antibody [E47] (ab32509)

All lanes : Anti-Mutant p53 antibody [E47] (ab32509) at 1/200 dilution

Lane 1 : A549 (Human lung carcinoma epithelial cell), Whole cell lysate
Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell), Whole cell lysate
Lane 3 : MCF-7 (Human breast adenocarcinoma epithelial cell), Whole cell lysate
Lane 4 : T-47D (Human ductal breast epithelial tumor epithelial cell), Whole cell lysate
Lane 5 : A431 (Human epidermoid carcinoma epithelial cell), Whole cell lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 44 kDa

Blocking/Diluting buffer and concentration: 5% NFDM/TBST

Exposure time: 180 seconds

Lane 1-3: Wildtype p53 cell lines

Lane 4-5: Mutant p53 cell lines

Observed MW: 50kDa, 39kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [E47] (ab32509)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma sections labeling mutant p53 with ab32509 at 1/2000 dilution (0.49 μg/mL). Sections were counterstained with Hematoxylin. Goat Anti-Rabbit IgG H&L (HRP Polymer) was used as the secondary antibody. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

Nuclear staining on human lung carcinoma.
Immunocytochemistry/ Immunofluorescence - Anti-Mutant p53 antibody [E47] (ab32509)

Immunocytochemistry/Immunofluorescence analysis of A431 (human epidermoid carcinoma) labelling mutant p53 with purified ab32509 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (ab150077). Nuclei counterstained with DAPI (blue).

Control: PBS only.
Immunoprecipitation - Anti-Mutant p53 antibody [E47] (ab32509)

Purified ab32509 at 1/50 dilution (2µg) immunoprecipitating Mutant p53 in HEK-293 whole cell lysate.
Lane 1 (input): HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab32509 + HEK-293 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32509 in HEK-293 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 53 kDa
Flow Cytometry - Anti-Mutant p53 antibody [E47] (ab32509)

Flow Cytometry analysis of HEK-293 (Human embryonic kidney epithelial cell) cells labeling Mutant p53 with purified ab32509 at 1/100 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cells without incubation with primary antibody and secondary antibody (Blue).
Western blot - Anti-Mutant p53 antibody [E47] (ab32509)

Anti-Mutant p53 antibody [E47] (ab32509) at 1/1000 dilution + A431 lysate

Predicted band size: 44 kDa
Observed band size: 53 kDa
why is the actual band size different from the predicted?
Anti-Mutant p53 antibody [E47] (ab32509)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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