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Product Info Summary
| SKU: | PB9330 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Calpain 1/CAPN1 Antibody Picoband®
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. 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/CAPN1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9330)
Host
Rabbit
Contents
Each vial contains antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3.
*This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
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.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9330 is reactive to CAPN1 in Human, Mouse, Rat
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
PB9330 is guaranteed for Flow Cytometry, IF, IHC, 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, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Immunocytochemistry/Immunofluorescence, 2μg/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, human T47D whole cell, human A549 whole cell, rat testis tissue, mouse lung tissue
IHC: Human Mammary Cancer tissue
ICC/IF: U20S cell
FCM: PC-3 cell
Validation Images & Assay Conditions
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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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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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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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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 fixed with 4% paraformaldehyde and 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.
Specific Publications For Anti-Calpain 1/CAPN1 Antibody Picoband® (PB9330)
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4 Customer Q&As for Anti-Calpain 1/CAPN1 Antibody Picoband®
Question
We are currently using anti-Calpain 1/CAPN1 antibody PB9330 for mouse tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on canine tissues as well?
Verified Customer
Verified customer
Asked: 2019-09-16
Answer
The anti-Calpain 1/CAPN1 antibody (PB9330) has not been validated for cross reactivity specifically with canine tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in canine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-09-16
Question
We have been able to see staining in mouse pancreas placenta. Do you have any suggestions? Is anti-Calpain 1/CAPN1 antibody supposed to stain pancreas placenta positively?
Verified Customer
Verified customer
Asked: 2018-08-31
Answer
From what I have seen in literature pancreas placenta does express CAPN1. From what I have seen in Uniprot.org, CAPN1 is expressed in esophagus mucosa, kidney, pancreas placenta, erythroleukemia, liver, among other tissues. Regarding which tissues have CAPN1 expression, here are a few articles citing expression in various tissues:
Erythroleukemia, Pubmed ID: 23186163
Kidney, Pancreas, and Placenta, Pubmed ID: 15489334
Liver, Pubmed ID: 24275569
Boster Scientific Support
Answered: 2018-08-31
Question
My team were satisfied with the WB result of your anti-Calpain 1/CAPN1 antibody. However we have seen positive staining in esophagus mucosa cytoplasm using this antibody. Is that expected? Could you tell me where is CAPN1 supposed to be expressed?
Verified Customer
Verified customer
Asked: 2018-07-11
Answer
According to literature, esophagus mucosa does express CAPN1. Generally CAPN1 expresses in cytoplasm. Regarding which tissues have CAPN1 expression, here are a few articles citing expression in various tissues:
Erythroleukemia, Pubmed ID: 23186163
Kidney, Pancreas, and Placenta, Pubmed ID: 15489334
Liver, Pubmed ID: 24275569
Boster Scientific Support
Answered: 2018-07-11
Question
I would like using your anti-Calpain 1/CAPN1 antibody for positive regulation of cell population proliferation studies. Has this antibody been tested with western blotting on jurkat whole cell lysate? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2018-04-05
Answer
Thank you for your inquiry. This PB9330 anti-Calpain 1/CAPN1 antibody is tested on rat lung tissue, tissue lysate, mouse lung tissue, a549 whole cell lysate, colo320 whole cell lysate, jurkat whole cell lysate, mammary cancer tissue. It is guaranteed to work for WB in human, mouse, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2018-04-05
