PA5-32349

antibody from Invitrogen Antibodies

Targeting: CHGA

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Recommended by provider)

Immunohistochemistry (Data presented on provider website)

Antibody Data

Product number

PA5-32349

Provider

Invitrogen Antibodies

Product name

Anti-Chromogranin A Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 10 min at room temperature. A suggested positive control is pancreas tissue.

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

500 µL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

N-Myc promotes therapeutic resistance development of neuroendocrine prostate cancer by differentially regulating miR-421/ATM pathway.

Yin Y, Xu L, Chang Y, Zeng T, Chen X, Wang A, Groth J, Foo WC, Liang C, Hu H, Huang J
Molecular cancer 2019 Jan 18;18(1):11

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The Polycomb-Dependent Epigenome Controls β Cell Dysfunction, Dedifferentiation, and Diabetes.

Lu TT, Heyne S, Dror E, Casas E, Leonhardt L, Boenke T, Yang CH, Sagar, Arrigoni L, Dalgaard K, Teperino R, Enders L, Selvaraj M, Ruf M, Raja SJ, Xie H, Boenisch U, Orkin SH, Lynn FC, Hoffman BG, Grün D, Vavouri T, Lempradl AM, Pospisilik JA
Cell metabolism 2018 Jun 5;27(6):1294-1308.e7

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of Chromogranin A was achieved by transfecting SHSY5Y cells with Chromogranin A specific siRNAs (Silencer® select Product # s2992, Product # 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 (Product # PA5-32349, 1:500 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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 Polyclonal Antibody (Product # PA5-32349) and a ~60 kDa band corresponding to Chromogranin A was observed in SH-SY5Y and IMR-32 cell lines treated with Protein Transport Inhibitor (PTI, 1X for 4hr) and FL1-06 (10uM for 4hr). 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), IMR-32 (Lane 4), IMR-32 treated with PTI (1X for 4hr) (Lane 5) and IMR-32 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 (1:500 dilution) 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).