Anti-Calpain 2 antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 2 antibody (AB39165)

ab39165 (4μg/ml) staining CALPAIN2 in human lung using an automated system (DAKO Autostainer Plus). Using this protocol there is cytoplasmic staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

• WB

Collaborator

Western blot - Anti-Calpain 2 antibody (AB39165)

ab39165 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 ab39165 overnight at 4 °C at a 1/2000 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 (ab39165)

Predicted band size: 80 kDa

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• WB

Unknown

Western blot - Anti-Calpain 2 antibody (AB39165)

All lanes:

Western blot - Anti-Calpain 2 antibody (ab39165) at 1/1000 dilution

Lane 1:

Calpain 2 control

Lane 2:

Cell lysates from human chondrosarcoma (treated with IL1 alpha) at 10 µg

Predicted band size: 80 kDa

Observed band size: 76 kDa

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