Antibodies, Proteins, Kits and Reagents for Life Science

358 492 ₸ Product size

100 µl 358 492 ₸ 10 µl 144 067 ₸

Order now and get it on Tuesday March 02, 2021

Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP503Y] to SRC Family (phospho Y418)
Suitable for: WB, Flow Cyt, ICC/IF, ELISA
Reacts with: Rat, Human

Product name

Anti-SRC Family (phospho Y418) antibody [EP503Y]
See all SRC Family primary antibodies
Description

Rabbit monoclonal [EP503Y] to SRC Family (phospho Y418)
Host species

Rabbit
Specificity

ab40660 recognises Lyn phosphorylation on Tyrosine 396. BLAST analysis show 100% homology with Hck (pY410) and Lck (pY393), and 92% homology with Src (pY418), Fyn (pY419) and Yes (pY425) Therefore, the antibody may cross-react with these proteins.

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Rat Human
WB	Rat Human

See all applications and species data

Immunogen

Synthetic peptide within Human SRC Family aa 350-450 (phospho Y418). The exact sequence is proprietary.
Database link: P07948
(Peptide available as ab202645)
Positive control
- WB: HeLa and C6 cell lysates treated with pervanadate. ICC/IF: HeLa cells treated with pervanadate. Flow Cyt: HeLa cells treated with pervanadate.
General notes
This protein is known to be similar in amino acid sequence to HCK (P08631), LCK (P06239), FYN (P06241), YES1 (P07947), and SRC (P12931). Therefore, cross-reactivity with these homologous proteins may be observed. We would be happy to provide immunogen alignment information upon request.
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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

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

Protein A purified
Clonality

Monoclonal
Clone number

EP503Y
Isotype

IgG
Research areas

Immunocytochemistry/ Immunofluorescence - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)

Immunocytochemistry/Immunofluorescence analysis of HeLa cells (untreated and treated with 1mM pervanadate for 15 minutes) labelling SRC Family (phospho Y418) with purified ab40660 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/250) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).
Flow Cytometry - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)

Flow Cytometry analysis of HeLa cells (treated with 1mM pervanadate for 30 minutes) labelling SRC Family (phospho Y418) with purified ab40660 at 1/80 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody.
Black - Isotype control, rabbit monoclonal IgG with pervanade treated HeLa cells.
Blue - Unlabelled control, pervanadate treated HeLa cells without incubation with primary and secondary antibodies.
Red - Pervandate treated HeLa cells with ab40660.
Green - Untreated HeLa cells labelled with ab40660.
Western blot - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)

All lanes : Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660) at 1/2000 dilution (purified)

Lane 1 : Untreated HeLa whole cell lysate
Lane 2 : Pervanadate treated HeLa whole cell lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 61 kDa
Observed band size: 56-59 kDa
why is the actual band size different from the predicted?

Exposure time: 3 seconds

Blocking and dilution buffer: 5% BSA/TBST.
Western blot - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)

All lanes : Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660) at 1/1000 dilution (purified)

Lane 1 : Pervanadate treated Jurkat cell lysate with Lyn non-phospho peptide
Lane 2 : Pervanadate treated Jurkat cell lysate with Lyn (pY396) phospho peptide
Lane 3 : Pervanadate treated Jurkat cell lysate with Lck non-phospho peptide
Lane 4 : Pervanadate treated Jurkat cell lysate with Lck (pY393) phospho peptide
Lane 5 : Pervanadate treated Jurkat cell lysate with Src non-phospho peptide
Lane 6 : Pervanadate treated Jurkat cell lysate with Src (pY418) phospho peptide

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

Predicted band size: 61 kDa
Observed band size: 56-59 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)

All lanes : Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660) at 1/1000 dilution (purified)

Lane 1 : Untreated C6 whole cell lysate
Lane 2 : Pervanadate treated C6 whole cell lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 61 kDa
Observed band size: 56-59 kDa why is the actual band size different from the predicted?

Exposure time: 1 second

Blocking and dilution buffer: 5% BSA/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)

ICC/IF image of unpurified ab40660 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab40660, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor^® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ELISA - Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)

Direct ELISA antibody dose-response curve using unpurified ab40660 at 0-1000 ng/ml. Antigen concentration of 1000 ng/mL. An alkaline phosphatase-conjugated goat anti-rabbit IgG (H+L) (1/2500) was used as the secondary antibody.
Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660)

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