Antibodies, Proteins, Kits and Reagents for Life Science

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [Y497] to Lyn
Suitable for: WB
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-Lyn antibody [Y497]
See all Lyn primary antibodies
Description

Rabbit monoclonal [Y497] to Lyn
Host species

Rabbit
Tested Applications & Species

Application Species
WB
Mouse

Rat

Human

See all applications and species data
Immunogen

Synthetic peptide within Human Lyn aa 1-100. The exact sequence is proprietary.
Database link: P07948
Epitope

Residues close to SH3 domain of human Lyn
Positive control
- WB: HAP1, Ramos, PC-12, and A431 whole cell lysates. Mouse lung, and mouse and rat liver tissue lysates.
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Application	Species
WB	Mouse Rat 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

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

Y497
Isotype

IgG
Research areas

Western blot - Anti-Lyn antibody [Y497] (ab32398)

All lanes : Anti-Lyn antibody [Y497] (ab32398) at 1/1000 dilution (Purified)

Lane 1 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 2 : Mouse lung lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : Rat liver lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 53 kDa
Observed band size: 56 kDa
why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Western blot - Anti-Lyn antibody [Y497] (ab32398)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Lyn knockout HAP1 cell lysate (20 µg)
Lane 3: Ramos cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab32398 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab32398 was shown to specifically react with Lyn when Lyn knockout samples were used. Wild-type and Lyn knockout samples were subjected to SDS-PAGE. ab32398 and ab8245 (loading control to GAPDH) were diluted at 1/2000 and 1/10 000 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.
Western blot - Anti-Lyn antibody [Y497] (ab32398)

All lanes : Anti-Lyn antibody [Y497] (ab32398) at 1/2000 dilution

Lane 1 : A431 whole cell lysate
Lane 2 : Mouse liver tissue lysate
Lane 3 : Rat liver tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 53 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?

Lanes 1 - 3: Merged signal (red and green). Green - ab32398 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab32398 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.

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