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Anti-Calpain 1 Antibody Picoband™ (monoclonal, 2I10)

     
  •  - Anti-Calpain 1 Antibody Picoband™ (monoclonal, 2I10) ABO15062
    Figure 1. Western blot analysis of Calpain 1 using anti-Calpain 1 antibody (M01943-3).
    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 lysates,
    Lane 2: human Hela whole cell lysates,
    Lane 3: human K562 whole cell lysates,
    Lane 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 # M01943-3) at 0.5 µg/mL overnight at 4°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 # EK1001) 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.
    detail
  •  - Anti-Calpain 1 Antibody Picoband™ (monoclonal, 2I10) ABO15062
    Figure 2. IF analysis of Calpain 1 using anti-Calpain 1 antibody (M01943-3).
    Calpain 1 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL mouse anti-Calpain 1 Antibody (M01943-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
    detail
  •  - Anti-Calpain 1 Antibody Picoband™ (monoclonal, 2I10) ABO15062
    Figure 3. Flow Cytometry analysis of A549 cells using anti-Calpain 1 antibody (M01943-3).
    Overlay histogram showing A549 cells stained with M01943-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Calpain 1 Antibody (M01943-3, 1 µg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immuno electron microscopy
  • EIA=Enzyme Immunoassay
WB, IF, ICC, FC
Primary Accession P07384
Host Mouse
Isotype Mouse IgG2a
Reactivity Human
Clonality Monoclonal
Format Lyophilized
Description Anti-Calpain 1 Antibody Picoband™ (monoclonal, 2I10) . Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human.
Reconstitution Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Additional Information
Gene ID 823
Other Names Calpain-1 catalytic subunit, 3.4.22.52, Calcium-activated neutral proteinase 1, CANP 1, Calpain mu-type, Calpain-1 large subunit, Cell proliferation-inducing gene 30 protein {ECO:0000303|Ref.3}, Micromolar-calpain, muCANP, CAPN1 (HGNC:1476), CANPL1
Calculated MW 82 kDa
Application Details Western blot, 0.25-0.5 µg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry, 1-3 µg/1x10^6 cells, Human
Contents Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clone Names Clone: 2I10
Immunogen E.coli-derived human Calpain 1 recombinant protein (Position: Q396-A555). Human Calpain 1 shares 86% amino acid (aa) sequence identity with both mouse and rat Calpain 1.
Purification Immunogen affinity purified.
Storage Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Protein Information
Name CAPN1 (HGNC:1476)
Synonyms CANPL1
Function Calcium-regulated non-lysosomal thiol-protease which catalyzes limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction (PubMed:19617626, PubMed:21531719, PubMed:2400579). Proteolytically cleaves CTBP1 at 'Asn-375', 'Gly-387' and 'His-409' (PubMed:23707407). Cleaves and activates caspase-7 (CASP7) (PubMed:19617626).
Cellular Location Cytoplasm. Cell membrane. Note=Translocates to the plasma membrane upon Ca(2+) binding. In granular keratinocytes and in lower corneocytes, colocalizes with FLG and FLG2 (PubMed:21531719)
Tissue Location Ubiquitous.
Research Areas
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Background

CAPN1 is also known as CANP or muCL. 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.

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$ 370.00
Cat# ABO15062
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