Anti-Calpain 1/CAPN1 Antibody Picoband™

Product Name

Anti-Calpain 1/CAPN1 Antibody Picoband™

View all Calpain 1 Antibodies

SKU/Catalog Number

PB9330

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Calpain 1/CAPN1 Antibody Picoband™ catalog # PB9330. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

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/CAPN1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9330)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

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.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PB9330 is reactive to CAPN1 in Human, Mouse, Rat

Applications

PB9330 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

82 kDa

Calculated molecular weight

81.89kDa

Background of Calpain 1

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.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

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, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of Calpain 1 using anti-Calpain 1 antibody (PB9330).
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 30 ug 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,
Lane 5: human T47D whole cell lysates,
Lane 6: human A549 whole cell lysates,
Lane 7: rat testis tissue lysates,
Lane 8: mouse lung tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Calpain 1 antigen affinity purified polyclonal antibody (Catalog # PB9330) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Calpain 1 at approximately 82 kDa. The expected band size for Calpain 1 is at 82 kDa.

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Figure 2. IHC analysis of Calpain 1 using anti-Calpain 1 antibody (PB9330).
Calpain 1 was detected in a paraffin-embedded section of Human Mammary Cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Calpain 1 Antibody (PB9330) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IF analysis of Calpain 1 using anti-Calpain 1 antibody (PB9330).
Calpain 1 was detected in immunocytochemical section of U20S 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 2μg/mL rabbit anti-Calpain 1 Antibody (PB9330) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) 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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Figure 4. Flow Cytometry analysis of PC-3 cells using anti-Calpain 1 antibody (PB9330).
Overlay histogram showing PC-3 cells stained with PB9330 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Calpain 1 Antibody (PB9330,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For CAPN1 (Source: Uniprot.org, NCBI)

Alternative Names

Calcium-activated neutral proteinase 1; calpain 1, (mu/I) large subunit; Calpain mu-type; calpain, large polypeptide L1; calpain-1 catalytic subunit; Calpain-1 large subunit; CANP 1; CANP; CANPL1EC 3.4.22.52; Cell proliferation-inducing gene 30 protein; cell proliferation-inducing protein 30; EC 3.4.22; micromolar-calpain; muCANPCANP1; muCL CAPN1 CANP, CANP1, CANPL1, SPG76, muCANP, muCL calpain 1 calpain-1 catalytic subunit|CANP 1|calcium-activated neutral proteinase 1|calpain 1, (mu/I) large subunit|calpain mu-type|calpain, large polypeptide L1|calpain-1 large subunit|cell proliferation-inducing gene 30 protein|cell proliferation-inducing protein 30|micromolar-calpain

For more info on CAPN1, check out the CAPN1 Infographic

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In this infographic, you will see the following information for CAPN1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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