Anti-Lyn (phospho Y397) 抗体 [EPR26354-1] (BSA and Azide free) (ab300119)

This data was developed using ab300118, the same antibody clone in a different buffer formulation. 

Blocking and dilution buffer and concentration: 5% NFDM/TBST. 

Dasatinib treatment inhibits the phosphorylation of Lyn at Y397 (PMID: 18451159).
The antibody may cross-react with phosphorylated HCK(Tyr411), LCK(Tyr394), BLK(Tyr389), Src(Tyr416), Fyn(Tyr420) and Yes(Tyr426).

This data was developed using ab300118, the same antibody clone in a different buffer formulation. 

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

This data was developed using ab300118, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized COLO 201 (human derived from metastatic site ascites bipolar, slightly refractile, fibroblast-like cell) cells lebelling Lyn (phospho Y397) with ab300118 at 1/50 (9.9 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2µg/ml) dilution (Green). Confocal image showing membrane staining in COLO 201 cells. The signal decreases after treatment with 200 mM Dasatinib for 24 hours. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5 µg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: PBS was used instead of ab300118 following by secondary antibody (ab150081) Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/ml) dilution.

This data was developed using ab300118, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed and 90% methanol parmeabilisied COLO 201 (Human colon adenocarcinoma) 200 nM Dasatinib for 24 hours (Red) / Untreated control (Green) cells labelling Lyn (phospho Y397) with ab300118 at 1/500 dilution (0.1µg) (Red) and Green compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Dasatinib treatment inhibit the phosphorylation of Lyn at Y397 (PMID: 18451159).

This data was developed using ab300118, the same antibody clone in a different buffer formulation.

Lyn (phospho Y397) was immunoprecipitated from 0.35 mg COLO 205 (human colorectal adenocarcinoma cell) whole cell lysate 10 µg with ab300118 at 1/30 dilution (2µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab300118 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 (Input): COLO 205 (human colorectal adenocarcinoma cell) whole cell lysate 10 µg

Lane 2 (+): COLO 205 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300118 in COLO 205 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 15 seconds

This data was developed using ab300118, the same antibody clone in a different buffer formulation.

Dot blot analysis of Lyn (phospho Y397) using ab300118 at 1:1000 (0.495 µg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjµgated (ab97051) at 1:100,000 dilution.

Lane 1: Human Lyn (phospho Y397) peptide a 

Lane 2: Human Lyn (phospho Y397) peptide b

Lane 3: Human Lyn non-phospho peptide

Exposure time: 3 minutes

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