Recombinant Anti-Calpain 2 antibody [EPR2562Y] (ab75994)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2562Y] to Calpain 2
- Suitable for: Flow Cyt (Intra), WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Calpain 2 antibody [EPR2562Y]
See all Calpain 2 primary antibodies -
Description
Rabbit monoclonal [EPR2562Y] to Calpain 2 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WBmore details
Unsuitable for: ICC/IF or IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Calpain 2 aa 150-250. The exact sequence is proprietary.
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Positive control
- WB: HEK-293T, A431 and LNCaP cell lysates; HeLa cytoplasmic or membrane lysate. Flow Cyt (intra): Permeabilized HeLa cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2562Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab75994 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/60.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/2000. Detects a band of approximately 80 kDa.
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Notes |
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Flow Cyt (Intra)
1/60. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/2000. Detects a band of approximately 80 kDa. |
Target
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Function
Calcium-regulated non-lysosomal thiol-protease which catalyze limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction. -
Tissue specificity
Ubiquitous. -
Sequence similarities
Belongs to the peptidase C2 family.
Contains 1 calpain catalytic domain.
Contains 3 EF-hand domains. -
Cellular localization
Cytoplasm. Cell membrane. Translocates to the plasma membrane upon Ca(2+) binding. - Information by UniProt
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Database links
- Entrez Gene: 824 Human
- Omim: 114230 Human
- SwissProt: P17655 Human
- Unigene: 350899 Human
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Alternative names
- Calcium activated neutral proteinase 2 antibody
- Calcium activated neutral proteinase antibody
- Calcium-activated neutral protease 2, catalytic subunit antibody
see all
Images
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ab75994 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 ab75994 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.
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All lanes : Anti-Calpain 2 antibody [EPR2562Y] (ab75994) at 1/2000 dilution
Lane 1 : Wild-type HEK-293T cell lysate at 40 µg
Lane 2 : CAPN2 knockout HEK-293T cell lysate at 40 µg
Lane 3 : A431 cell lysate at 20 µg
Lane 4 : LNCaP cell lysate at 20 µg
Performed under reducing conditions.
Observed band size: 75 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab75994 observed at 75 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab75994 was shown to react with Calpain 2 in wild-type HEK-293T cells in Western blot with loss of signal observed in CAPN2 knockout cell line ab266628 (CAPN2 knockout cell lysate ab257379). Wild-type HEK-293T and CAPN2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab75994 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Intracellular flow cytometric analysis of permeabilized HeLa cells using ab75994 (red) or a rabbit IgG (negative) (green).
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Anti-Calpain 2 antibody [EPR2562Y] (ab75994) at 1/500 dilution + HeLa cell lysate at 10 µg
Secondary
HRP labelled goat anti-rabbit at 1/2000 dilution
Observed band size: 75 kDa why is the actual band size different from the predicted?
Datasheets and documents
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SDS download
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Datasheet download
References (2)
ab75994 has been referenced in 2 publications.
- Xu F et al. Calpain-2 Enhances Non-Small Cell Lung Cancer Progression and Chemoresistance to Paclitaxel via EGFR-pAKT Pathway. Int J Biol Sci 15:127-137 (2019). PubMed: 30662353
- Sun L et al. GAS2-Calpain2 axis contributes to the growth of leukemic cells. Acta Biochim Biophys Sin (Shanghai) 47:795-804 (2015). PubMed: 26358320