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Product Info Summary
| SKU: | M01943-3 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Mouse |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-Calpain 1 Antibody Picoband® (monoclonal, 2I10)
SKU/Catalog Number
M01943-3
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Calpain 1 Antibody Picoband® (monoclonal, 2I10) catalog # M01943-3. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
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.
Cite This Product
Anti-Calpain 1 Antibody Picoband® (monoclonal, 2I10) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M01943-3)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Monoclonal
Clone Number
2I10
Isotype
Mouse IgG2a
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.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M01943-3 is reactive to CAPN1 in Human
Observed Molecular Weight
82 kDa
Calculated molecular weight
81.9 kDa
Background of CAPN1
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.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
M01943-3 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human Jurkat whole cell, human Hela whole cell, human K562 whole cell, human PC-3 whole cell
ICC/IF: A431 cell
FCM: A549 cell
Validation Images & Assay Conditions
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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.
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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 5μg/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.
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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 fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with mouse anti-Calpain 1 Antibody (M01943-3, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-Calpain 1 Antibody Picoband® (monoclonal, 2I10) (M01943-3)
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