MA1-41203

antibody from Invitrogen Antibodies

Targeting: LCK

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-41203

Provider

Invitrogen Antibodies

Product name

LCK Monoclonal Antibody (33D196)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

The immunogen sequence is 100% homologus in mouse. Suggested positive control: Jurkat whole cell lysate.

Reactivity

Human, Mouse

Host

Mouse

Isotype

IgG

Antibody clone number

33D196

Vial size

200 μL

Concentration

1.0 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of LCK in 20 µg/lane Jurkat cell lysate. Samples were incubated in LCK monoclonal antibody (Product # MA1-41203) using a dilution of 1:1000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-LCK Monoclonal Antibody (33D196) (Product # MA1-41203) and a 54kDa band corresponding to LCK was observed in Jurkat, MOLT-4 and A549, but not in Ramos, SK-BR-3 and SK-OV-3. An uncharacterized band was alos observed at ~ 65kDa in few cell lines. Whole cell lysates (30ug lysate) of Jurkat (Lane 1), Ramos (Lane 2), MOLT-4 (Lane 3), A549 (Lane 4), SK-BR-3 (Lane 5) and SK-OV-3 (Lane 6) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The bot was probed with the primary antibody (1:1000 dilution) detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of LCK was achieved by transfecting Jurkat with LCK specific siRNAs (Silencer® select Products # s8108, s8107). Western blot analysis (Fig. a) was performed using whole cell extracts from the LCK knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with LCK Monoclonal Antibody (33D196) (Product # MA1-41203, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to LCK. An uncharacterized band was observed at ~60kDa.