LF-MA0140

antibody from Invitrogen Antibodies

Targeting: CHGA

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

LF-MA0140

Provider

Invitrogen Antibodies

Product name

Anti-Chromogranin A Monoclonal Antibody (23A1)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

A suggested positive control for this product is Bosc23 cells transfected with myc/His-CGA.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

23A1

Vial size

100 µL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of Chromogranin A was achieved by transfecting SH-SY5Y cells with Chromogranin A specific siRNAs (Silencer® select Product # s2992, s2993) and further treating with PTI (1X for 4hr). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from Chromogranin A knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Chromogranin A Monoclonal Antibody (23A1) (Product # LF-MA0140, 1:2000 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 Chromogranin A.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Chromogranin A Monoclonal Antibody (Product # LF-MA0140) and a ~60 kDa band corresponding to Chromogranin A was observed in SH-SY5Y cell line treated with Protein Transport Inhibitor (PTI, 1X for 4r) and FL1-06 (10uM for 4hr) as compared to Hep G2 cell line treated with same inhibitors. Whole cell extracts (30ug) of SH-SY5Y (Lane 1), SH-SY5Y treated with PTI (1X for 4hr) (Lane 2), SH-SY5Y treated with FL1-06 (10uM for 4hr) (Lane 3), Hep G2 (Lane 4), Hep G2 treated with PTI (1X for 4hr) (Lane 5) and Hep G2 treated with FL1-06 (10uM for 4hr) (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 blot was probed with the primary antibody (1:2000 dilution) and 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).