Anti-Calpain 2 antibody [EPR5977]

• WB

Collaborator

Western blot - Anti-Calpain 2 antibody [EPR5977] (AB126600)

ab126600 was shown to react with CAPN2 in wild-type MDA-MB-231 (wpxld GFP) cells in Western blot with loss of signal observed in a CAPN2 knockout cell line. Wild-type MDA-MB-231 (wpxld GFP) and CAPN2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab126600 overnight at 4 °C at a 1/5000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2 μg/ml before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR5977] (ab126600)

Predicted band size: 80 kDa

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• WB

Collaborator

Western blot - Anti-Calpain 2 antibody [EPR5977] (AB126600)

ab126600 was shown to react with CAPN2 in wild-type MDA-MB-231 (wpxld GFP) and HEK293T cells in Western blot with loss of signal observed in a CAPN2 knockout cell lines. Wild-type MDA-MB-231 (wpxld GFP) and HEK293T ab255449 (CAPN2 knockout cell lysate ab257379) and CAPN2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab126600 overnight at 4 °C at a 1/5000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2 μg/ml before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR5977] (ab126600)

Predicted band size: 80 kDa

false

• WB

Lab

Western blot - Anti-Calpain 2 antibody [EPR5977] (AB126600)

Lanes 1-4 : Merged signal (red and green). Green - ab126600 observed at 80 kDa. Red - loading control ab8245 observed at 36 kDa.

ab126600 Anti-Calpain 2 antibody [EPR5977] was shown to specifically react with Calpain 2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266628 (knockout cell lysate ab257379) was used. Wild-type and Calpain 2 knockout samples were subjected to SDS-PAGE. ab126600 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR5977] (ab126600) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

CAPN2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human CAPN2 (Calpain 2) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-capn2-calpain-2-knockout-hek-293t-cell-line-ab266628'>ab266628</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

LNCaP cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 80 kDa

Observed band size: 80 kDa

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• WB

Unknown

Western blot - Anti-Calpain 2 antibody [EPR5977] (AB126600)

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR5977] (ab126600) at 1/1000 dilution

Lane 1:

Rat brain lysates at 20 µg

Lane 2:

Rat kidney lysates at 20 µg

Lane 3:

PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 4:

Mouse brain lysates at 20 µg

Lane 5:

Mouse kidney lysates at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 80 kDa

Observed band size: 80 kDa

false

• WB

Unknown

Western blot - Anti-Calpain 2 antibody [EPR5977] (AB126600)

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR5977] (ab126600) at 1/1000 dilution

Lane 1:

PC3 cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

A431 cell lysate at 10 µg

Lane 4:

293T cell lysate at 10 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/2000 dilution

Predicted band size: 80 kDa

Observed band size: 80 kDa

false

• WB

CiteAb

Western blot - Anti-Calpain 2 antibody [EPR5977] (AB126600)

Calpain 2 western blot using anti-Calpain 2 antibody [EPR5977] ab126600. Publication image and figure legend from Zeng, X., Chen, S., et al., 2017, Oncotarget, PubMed 29069832.

ab126600 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab126600 please see the product overview.

AG/UnAG attenuates calpain system activity in co-cultured myotubes(A) Western blot of μ-calpain, m-calpain, calpain-3, calpastatin and GAPDH in C2C12 myotubes. (B) Quantification of μ-calpain was normalized to GAPDH. Significant differences were detected between CO and any NC groups (#p < 0.001), between CO and CO+AG/UnAG groups (*p < 0.001), by one-way ANOVA followed by Tukey test. (C-E) Quantification of m-calpain, calpain-3 and calpastatin was normalized to GAPDH. No significant differences were detected among the six groups, by one-way ANOVA followed by Tukey test. (F) The levels of μ-calpain mRNA in C2C12 myotubes. mRNA levels were normalized to GAPDH. Significant differences were detected between NC and CO groups (#P = 0.005, P = 0.007; P = 0.007 respectively), between CO and CO+AG groups (*P = 0.01, P = 0.015 respectively), by one-way ANOVA followed by Tukey test. Data are represented as mean ± SD.

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