Anti-c-Kit (phospho Y823)抗体

• ICC

Supplier Data

Immunocytochemistry - Anti-c-Kit (phospho Y823) antibody (AB5634)

Jurkat cells stained for c-Kit cleave site (green) using ab5634 at 1/250 dilution in ICC/IF. It was followed by Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate at 1/2000 dilution for 45 minutes at room temperature (Panel a). Nuclei (Panel b : blue) were stained with SlowFade® Gold Antifade Mountant with DAPI. F-actin (Panel c : red) was stained with Rhodamine Phalloidin at 1/300 dilution. Panel d represents the merged image showing membranous localization. Panel e shows the untreated control cells with no signal. Panel f shows the no primary antibody control.

• WB

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Western blot - Anti-c-Kit (phospho Y823) antibody (AB5634)

Peptide Competition : Extracts prepared from M07e cells left untreated (1) or treated (2-5) with SCF were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with 0.50 μg/mL ab5634 antibody for two hours at room temperature in a 1% BSA-TBST buffer, following prior incubation with : no peptide (1, 2), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphotyrosine containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab5634 blocks the antibody signal, thereby demonstrating the specificity of the antibody. Peptide Competition : Extracts prepared from M07e cells left untreated (1) or treated (2-5) with SCF were resolved

All lanes:

Western blot - Anti-c-Kit (phospho Y823) antibody (ab5634)

Predicted band size: 109 kDa

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