PA5-44190

antibody from Invitrogen Antibodies

Targeting: GCLC GCS, GLCL, GLCLC

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-44190

Provider

Invitrogen Antibodies

Product name

GCLC Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Peptide sequence: RISKSRYDSI DSYLSKCGEK YNDIDLTIDK EIYEQLLQEG IDHLLAQHVA Sequence homology: Cow: 100%; Dog: 100%; Guinea Pig: 100%; Horse: 100%; Human: 100%; Mouse: 100%; Rabbit: 100%; Rat: 100%; Yeast: 100%; Zebrafish: 100%

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.5 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of human placenta cells using an anti-GCLC polyclonal antibody (Product # PA5-44190).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of GCLC was achieved by transfecting Hep G2 cells with GCLC specific siRNAs (Silencer® select Product # s5800). Western blot analysis (Fig. a) was performed whole cell extracts from the GCLC knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with GCLC Monoclonal Antibody (Product # PA5-44190, 1µg/ml) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25 µg/ml, 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 GCLC.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-GCLC Polyclonal Antibody (Product # PA5-44190) and a 72kDa band corresponding to GCLC was observed across cell lines and tissues tested. Whole cell extracts (30 ug lysate) of A549 (Lane 1), Hep G2 (Lane 2), MCF7 (Lane 3), Jurkat (Lane 4), Caco-2 (Lane 5), Mouse Liver (Lane 6) and Mouse Kidney (Lane 7) 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 blot was probed with the primary antibody (1µg/ml) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).