Western Blot (WB)
(Figure 1. Western blot analysis of Calpain 1 using anti-Calpain 1 antibody (MBS1754003).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: human Jurkat whole cell lysatesLane 2: human Hela whole cell lysatesLane 3: human K562 whole cell lysatesLane 4: human PC-3 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti- Calpain 1 antigen affinity purified monoclonal antibody (Catalog # MBS1754003) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for Calpain 1 at approximately 82KD. The expected band size for Calpain 1 is at 82KD. )
Mouse anti-Human Calpain 1 Monoclonal Antibody | anti-CAPN1 antibody
Anti-Calpain 1 Antibody (monoclonal, 2E3)
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of Calpain 1 using anti-Calpain 1 antibody (MBS1754003).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: human Jurkat whole cell lysatesLane 2: human Hela whole cell lysatesLane 3: human K562 whole cell lysatesLane 4: human PC-3 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti- Calpain 1 antigen affinity purified monoclonal antibody (Catalog # MBS1754003) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for Calpain 1 at approximately 82KD. The expected band size for Calpain 1 is at 82KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of Calpain 1 using anti-Calpain 1 antibody (MBS1754003).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: human Jurkat whole cell lysatesLane 2: human Hela whole cell lysatesLane 3: human K562 whole cell lysatesLane 4: human PC-3 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti- Calpain 1 antigen affinity purified monoclonal antibody (Catalog # MBS1754003) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for Calpain 1 at approximately 82KD. The expected band size for Calpain 1 is at 82KD. )
Flow Cytometry (FC/FACS)
(Figure 2. Flow Cytometry analysis of A549 cells using anti-Calpain 1 antibody (MBS1754003).Overlay histogram showing A549 cells stained with MBS1754003 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Calpain 1 Antibody (MBS1754003, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 2. Flow Cytometry analysis of A549 cells using anti-Calpain 1 antibody (MBS1754003).Overlay histogram showing A549 cells stained with MBS1754003 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Calpain 1 Antibody (MBS1754003, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Wang JC, et al. Proteolysis by endogenous calpain I leads to the activation of calcineurin in human heart. Clin Lab, 2012.
NCBI and Uniprot Product Information
NCBI Description
The calpains, calcium-activated neutral proteases, are nonlysosomal, intracellular cysteine proteases. The mammalian calpains include ubiquitous, stomach-specific, and muscle-specific proteins. The ubiquitous enzymes consist of heterodimers with distinct large, catalytic subunits associated with a common small, regulatory subunit. This gene encodes the large subunit of the ubiquitous enzyme, calpain 1. Several transcript variants encoding two different isoforms have been found for this gene. [provided by RefSeq, Nov 2010]
Uniprot Description
CAPN1: Calcium-regulated non-lysosomal thiol-protease which catalyze limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction. Forms a heterodimer with a small (regulatory) subunit (CAPNS1). Ubiquitous. Activated by micromolar concentrations of calcium and inhibited by calpastatin. Belongs to the peptidase C2 family.
Protein type: EC 3.4.22.52; Motility/polarity/chemotaxis; Protease
Chromosomal Location of Human Ortholog: 11q13
Cellular Component: focal adhesion; membrane; cytoplasm; plasma membrane; cytosol
Molecular Function: protein binding; calcium-dependent cysteine-type endopeptidase activity; calcium ion binding
Biological Process: extracellular matrix disassembly; extracellular matrix organization and biogenesis; receptor catabolic process; positive regulation of cell proliferation; proteolysis
Research Articles on CAPN1
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Product Notes
The CAPN1 capn1 (Catalog #AAA1754003) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-Calpain 1 Antibody (monoclonal, 2E3) reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's Calpain 1 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Flow Cytometry (FC/FACS/FCM). WB: 0.25-0.5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human|. Researchers should empirically determine the suitability of the CAPN1 capn1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Calpain 1, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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