Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸

Order now and get it on Wednesday March 10, 2021

Anti-Src (phospho Y529) antibody [Y232] (ab32078)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [Y232] to Src (phospho Y529)
Suitable for: WB, Dot blot, ICC/IF
Reacts with: Human

Product name

Anti-Src (phospho Y529) antibody [Y232]
See all Src primary antibodies
Description

Rabbit monoclonal [Y232] to Src (phospho Y529)
Host species

Rabbit
Specificity

ab32078 will detect Src phosphorylation on Tyrosine 529 of both isoforms. The antibody immunogen shares 93% homology with Fyn and Yes and 85% homology with Fgr. Therefore, it is likely that the antibody will cross-react with these proteins. However, this is just based on BLAST results and no experiments were performed. The sequence numbering is based off the mature form of the protein without the initiator methionine.
Tested Applications & Species

Application Species
ICC/IF
Human

WB
Human

See all applications and species data
Immunogen

Synthetic peptide within Human Src (phospho Y529). The exact sequence is proprietary. The sequence numbering is based off the mature form of the protein without the initiator methionine.
(Peptide available as ab179556)
Positive control
- WB: HeLa and A431 cell lysates ICC/IF: HeLa and A431 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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Application	Species
ICC/IF	Human
WB	Human

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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

Y232
Isotype

IgG
Research areas

Western blot - Anti-Src (phospho Y529) antibody [Y232] (ab32078)

WB analysis. Lane 1:Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates 15ug and Lane 2:HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with H₂O₂ at 10mM for 1 h. whole cell lysates 15ug and Lane 3:HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with H₂O₂ at 10mM for 1 h. whole cell lysates 15ug. Then the membrane was incubated with phosphatase. Primary ab used at 1:30,000 dilution. 5% NFDM/TBST was used as Blocking buffer and Diluting buffer. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as a Secondary ab at 1:20,000 dilution. The Exposure time was 30 seconds.
Immunocytochemistry/ Immunofluorescence - Anti-Src (phospho Y529) antibody [Y232] (ab32078)

Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labelling Src (phospho Y529) with ab32078 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] (1/200) - Microtubule Marker (Alexa Fluor^® 594). DAPI (blue) was used as a nuclear counterstain.

The green staining was increased in the H2O2 (10mM, 1 hour) treated HeLa cells when compared with HeLa cells without treatment. After LP treatment, the green signaling was obviously decreased.

For the pan antibody, there was no great difference after H₂O₂ (10 mM, 1 hour) or EGF (100 ng/mL, 10 minutes) + LP treatment.

The data showed mostly Cytoplasm and Membran staining for Ab32078.
Dot Blot - Anti-Src (phospho Y529) antibody [Y232] (ab32078)

Dot Blot analysis of Lane 1: Src (pY529) phospho peptide and Lane 2: Src non-phospho peptide labeling Src (phospho Y529) with ab32078 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. ab97051 Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 3 minutes.
Western blot - Anti-Src (phospho Y529) antibody [Y232] (ab32078)

All lanes : Anti-Src (phospho Y529) antibody [Y232] (ab32078) at 1/10000 dilution

Lane 1 : Untreated A431 cells
Lane 2 : EGF treated A431 cells
Lane 3 : EGF and alkaline phosphatase treated A431 cells

Predicted band size: 60 kDa
Observed band size: 55 kDa
why is the actual band size different from the predicted?
Dot Blot - Anti-Src (phospho Y529) antibody [Y232] (ab32078)

Dot Blot analysis on immunogen phospho peptide (A) and non phospho peptide (B) using 1/2000 ab32078.
Anti-Src (phospho Y529) antibody [Y232] (ab32078)

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