Antibodies, Proteins, Kits and Reagents for Life Science

358 492 ₸ Product size

100 µl 358 492 ₸

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 [Y5] to Mutant p53
Suitable for: ICC/IF, WB, IHC-P, Flow Cyt, IP
Reacts with: Human

Product name

Anti-Mutant p53 antibody [Y5]
See all Mutant p53 primary antibodies
Description

Rabbit monoclonal [Y5] to Mutant p53
Host species

Rabbit
Specificity

ab32049 recognises human mutant forms of p53 but not human p53 wild type. ab32049 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
WB	Human

See all applications and species data

Immunogen

Synthetic peptide within Human Mutant p53 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P04637
(Peptide available as ab204287)
General notes
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.
Dissociation constant (K_D)

K_D = 2.02 x 10 ^-10 M
Learn more about K_D
Storage buffer

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

IgG fraction
Clonality

Monoclonal
Clone number

Y5
Isotype

IgG
Research areas

Western blot - Anti-Mutant p53 antibody [Y5] (ab32049)

All lanes : Anti-Mutant p53 antibody [Y5] (ab32049) at 1/10000 dilution

Lane 1 : A431 (Human epidermoid carcinoma epithelial cell), Whole cell lysate
Lane 2 : A549 (Human lung carcinoma epithelial cell), Whole cell lysate
Lane 3 : MCF-7 (Human breast adenocarcinoma epithelial cell), Whole cell lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 44 kDa

Exposure time: 180 seconds

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

Lane1: Mutant p53 cell lines

Lane2-3: Wildtype p53 cell lines

Exposure time: 180 seconds

Observed MW: 50KDa
Western blot - Anti-Mutant p53 antibody [Y5] (ab32049)

All lanes : Anti-Mutant p53 antibody [Y5] (ab32049) at 1/500 dilution

Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell), Whole cell lysate
Lane 2 : A549(Human lung carcinoma epithelial cell), Whole cell lysate
Lane 3 : MCF-7(Human breast adenocarcinoma epithelial cell), Whole cell lysate
Lane 4 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell), Whole cell lysate
Lane 5 : MDA-MB-435(Human mammary gland ductal carcinoma melanocyte), Whole cell lysate
Lane 6 : T-47D (Human ductal breast epithelial tumor epithelial cell), Whole cell lysate
Lane 7 : Raji (Human Burkitt's lymphoma B lymphocyte), Whole cell lysate
Lane 8 : Ramos (Human Burkitt's lymphoma B lymphocyte), Whole cell lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 44 kDa

Exposure time: 60 seconds

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

Lane 1-4: Wildtype p53 cell lines

Lane 5-8: Mutant p53 cell lines

Observed MW: 50 kDa
Western blot - Anti-Mutant p53 antibody [Y5] (ab32049)

Anti-Mutant p53 antibody [Y5] (ab32049) at 1/500 dilution + A431 cell lysate

Predicted band size: 44 kDa
Observed band size: 50 kDa
why is the actual band size different from the predicted?
Immunocytochemistry/ Immunofluorescence - Anti-Mutant p53 antibody [Y5] (ab32049)

Confocal image showing nuclear staining on A431 cell line.

Immunocytochemistry/Immunofluorescence analysis of A431(Human epidermoid carcinoma cell line) labelling Mutant p53 with ab32049 at 1/100. Cells were fixed with 4% Paraformaldehyde (20 minutes) and permeabilized with 0.1% Triton X-100 (5 minutes). ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counterstained with ab195889, anti- alpha tubulin antibody (1/200) using an Alexa Fluor^® 594-conjugated microtubule marker as the secondary. Nuclei were counterstained with DAPI (blue).

Secondary Only Control: PBS was used instead of the primary antibody as the negative control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

ab32049 showing positive staining in Urinary bladder carcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cytometry - Anti-Mutant p53 antibody [Y5] (ab32049)

Flow Cytometry analysis of A431(human epidermoid carcinoma) cells labeling Mutant p53 with unpurified ab32049 at 1/20 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

Immunohistochemistry (Paraffin-embedded sections) using ab32049 at a dilution of 1/50 and human skin cancer

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

ab32049 showing positive staining in Glioma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

ab32049 showing positive staining in Gastric adenocarcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

ab32049 showing positive staining in Breast carcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

Immunohistochemical analysis of paraffin embedded normal Human breast tissue (negative control) labeling p53 with ab32049.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mutant p53 antibody [Y5] (ab32049)

Immunohistochemical analysis of paraffin embedded normal Human uterus tissue (negative control) labeling p53 with ab32049.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
OI-RD Scanning - Anti-Mutant p53 antibody [Y5] (ab32049)

Equilibrium disassociation constant (K_D)
Learn more about K_D

Click here to learn more about K_D
Anti-Mutant p53 antibody [Y5] (ab32049)

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