- PA5-17494 - Invitrogen Antibodies CAPN2 antibody

Submitted references Ca(2+) and calpain mediate capsaicin-induced ablation of axonal terminals expressing transient receptor potential vanilloid 1.

Wang S, Wang S, Asgar J, Joseph J, Ro JY, Wei F, Campbell JN, Chung MK
The Journal of biological chemistry 2017 May 19;292(20):8291-8303

Distinct roles for μ-calpain and m-calpain in synaptic NMDAR-mediated neuroprotection and extrasynaptic NMDAR-mediated neurodegeneration.

Wang Y, Briz V, Chishti A, Bi X, Baudry M
The Journal of neuroscience : the official journal of the Society for Neuroscience 2013 Nov 27;33(48):18880-92

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Calpain 2 Large Subunit (M-type) in extracts from C2C12 and PC3 cells using Calpain 2 Large Subunit (M-type) polyclonal antibody (Product # PA5-17494).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Calpain 2 (CAPN2) was achieved by transfecting A-431 with Calpain 2 (CAPN2) specific siRNAs (Silencer® select Products # s321). Western blot analysis (Fig. a) was performed using whole cell extracts from the Calpain 2 (CAPN2) knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Calpain 2 Polyclonal Antibody (Product # PA5-17494, 1:1000 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). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to CAPN2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Calpain 2 Polyclonal Antibody (Product # PA5-17494) and a 76 kDa band corresponding to Calpain 2 was observed in C2C12, DU 145, NIH/3T3, A-431 and PC-3, but not in Jurkat and Daudi which are reported to be negative. Whole cell lysates (30 µg lysate) of Jurkat (Lane 1), C2C12 (Lane 2), DU 145 (Lane 3), NIH/3T3 (Lane 4), A-431 (Lane 5), PC-3 (Lane 6) and Daudi (Lane 7) were electrophoresed using NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX). 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 (1:1000 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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of Calpain-2 was performed by loading 20 µg of WT (lane 1) and CAPN2 CRISPR KO (lane 2) MDA-MB-231 cell lysates in RIPA buffer onto a 4-15% gradient polyacrylamide gel. Proteins on the blots were visualized with Ponceau staining (below immunoblot). Proteins were transferred to nitrocellulose membrane and blocked in 5% milk for 1 hr. CAPN2 was detected at approximately 74 kDa using a CAPN2 polyclonal antibody (Product # PA5-17494) at a dilution of 1:1,000 in 5% BSA in TBS with 0.1% Tween 20 (TBST) overnight at 4°C. The peroxidase-conjugated secondary antibody (Product # 65-6120) was diluted to 0.2 µg/mL in TBST with 5% milk for 1 hr. Chemiluminescent detection was performed using Pierce ECL Western Blotting Substrate (Product # 32106). Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

PA5-17494

Antibody data