Anti-Calpain 1/CAPN1 Antibody


SKU PA1364
Size 100μg/vial
Reactivity Human, Mouse, Rat
Clonality Polyclonal
Host Rabbit
Ig Isotype N/A
Applications IHC, ICC, WB

Overview

Product Name Anti-Calpain 1/CAPN1 Antibody
SKU/Catalog Number PA1364
Storage & Handling At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.
Size 100μg/vial
Description Rabbit IgG polyclonal antibody for Calpain-1 catalytic subunit(CAPN1) detection. Tested with WB, IHC-P, IHC-F, ICC in Human;Mouse;Rat.
Cite This Product Anti-Calpain 1/CAPN1 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1364)
Host Rabbit
Contents/Buffer Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
Form Lyophilized
Immunogen A synthetic peptide corresponding to a sequence in the middle region of human Calpain 1(312-326aa EWNNVDPYERDQLRV), different from the mouse sequence by two amino acids.
Reactivity Human, Mouse, Rat

Assay Details

Assay Dilutions Overview

Concentration: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Immunocytochemistry , 0.5-1μg/ml, Human, -
Immunohistochemistry(Frozen Section), 0.5-1μg/ml, Rat, Human, Mouse
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat, By Heat
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat

Boster's Secondary Antibodies And IHC, WB Kits

The following reagents are used to generate the images below.

Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P), IHC(F) and ICC.

Images And Assay Conditions

Figure 2. IHC analysis of Calpain 1 using anti-Calpain 1 antibody (PA1364).
Calpain 1 was detected in paraffin-embedded section of rat lung tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Calpain 1 Antibody (PA1364) 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.

Figure 3. IHC analysis of Calpain 1 using anti-Calpain 1 antibody (PA1364).
Calpain 1 was detected in immunocytochemical section of HELA Cell. 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 1μg/ml rabbit anti-Calpain 1 Antibody (PA1364) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 4. IHC analysis of Calpain 1 using anti-Calpain 1 antibody (PA1364).
Calpain 1 was detected in paraffin-embedded section of human intestinal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Calpain 1 Antibody (PA1364) 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.

Figure 1. Western blot analysis of Calpain 1 using anti-Calpain 1 antibody (PA1364).
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 50ug of sample under reducing conditions.
Lane 1: mouse spleen tissue lysates,
Lane 2: mouse lung tissue lysates,
Lane 3: mouse kidney tissue lysates,
Lane 4: mouse brain tissue lysates,
Lane 5: mouse thymus tissue lysates,
Lane 6: mouse small intestine tissue lysates,
Lane 7: mouse ovary tissue 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 rabbit anti-Calpain 1 antigen affinity purified polyclonal antibody (Catalog # PA1364) 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 1:10000 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 80KD. The expected band size for Calpain 1 is at 80KD.

Figure 6. Western blot analysis of Calpain 1 using anti-Calpain 1 antibody (PA1364).
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 50ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: rat heart tissue lysates,
Lane 3: rat testis tissue lysates,
Lane 4: rat thymus tissue lysates,
Lane 5: human MM231 whole cell lysates,
Lane 6: human HeLa whole cell lysates,
Lane 7: human SMMC-7721 whole cell lysates,
Lane 8: human HT1080 whole cell lysates,
Lane 9: human Colo320 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 rabbit anti-Calpain 1 antigen affinity purified polyclonal antibody (Catalog # PA1364) 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 1:10000 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 80KD. The expected band size for Calpain 1 is at 80KD.

Figure 5. IHC analysis of Calpain 1 using anti-Calpain 1 antibody (PA1364).
Calpain 1 was detected in frozen section of rat cardiac muscle tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Calpain 1 Antibody (PA1364) 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.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id P07384
Gene Name CAPN1
Protein Name Calpain-1 catalytic subunit
Tissue Specificity Ubiquitous.
Alternative 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;Micromolar-calpain;muCANP;CAPN1;CANPL1;PIG30;
Subcellular Localization Cytoplasm . Cell membrane . Translocates to the plasma membrane upon Ca(2+) binding. In granular keratinocytes and in lower corneocytes, colocalizes with FLG and FLG2 (PubMed:21531719). .
Molecular Weight 81890 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein Function Calcium-regulated non-lysosomal thiol-protease which catalyze limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction.
Research Areas Human, Mouse, Rat

*You can search these to find other products in these research areas.
Background Calpain-1 catalytic subunit is a protein that in humans is encoded by the CAPN1 gene. Calpain is an intracellular protease that requires calcium for its catalytic activity. Two isozymes, calpain I(mu-calpain) and calpain II(m-calpain), with different calcium requirements, have been identified. Both are heterodimers composed of L(large, catalytic, 80 kD) and S(small, regulatory, 30 kD) subunits. The isozymes share an identical S subunit, with the differences arising from the L subunits, L1(CAPN1) and L2. By quantitative RT-PCR, Ueyama et al.(1998) found that expression of calpain-1 and calpain-2 mRNA was significantly increased in muscle biopsy samples derived from 5 men with progressive muscular dystrophy(e.g., DMD; 310200) and 2 men and 3 women with amyotrophic lateral sclerosis(ALS; 105400) compared with controls. Using cDNA clones as probes, Ohno et al.(1989, 1990) assign CANPL1 to chromosome 11.

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Polyclonal antibody for CALPAIN 1/CAPN1 detection. Host: Rabbit.Size: 100μg/vial. Tested applications: IHC-P. Reactive species: Human. CALPAIN 1/CAPN1 information: Molecular Weight: 81890 MW; Subcellular Localization: Cytoplasm . Cell membrane . Translocates to the plasma membrane upon Ca(2+) binding. In granular keratinocytes and in lower corneocytes, colocalizes with FLG and FLG2 (PubMed:21531719); Tissue Specificity: Ubiquitous.
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In stock
Order Product
PA1364
Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
*Sample sizes are prepared on demand and will take extra lead time. (cannot be conjugated)
$280.00

Troubleshooting

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Publications

Mo Xg, Chen Qw, Li Xs, Zheng Mm, Ke Dz, Deng W, Li Gq, Jiang J, Wu Zq, Wang L, Wang P, Yang Y, Cao Gy. Microvasc Res. 2011 Mar;81(2):160-8. Doi: 10.1016/J.Mvr.2010.12.004. Epub 2010 Dec 24. Suppression Of Nhe1 By Small Interfering Rna Inhibits Hif...
Chen Hx, Tang Sp, Gao Ft, Xu Jl, Jiang Xp, Cao J, Fu Gb, Sun K, Liu Sz, Shi W. Medicine (Baltimore). 2014 Nov;93(23):E138. Doi: 10.1097/Md.0000000000000138. Fibrosis, Adipogenesis, And Muscle Atrophy In Congenital Muscular Torticollis.

Customer Q&As

  • Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
    A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
  • Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WB
    A: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
  • Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
    A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
  • Q: What should I use for negative control?
    A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
  • Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
    A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
  • Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
    A: You can find the immunogen sequence under "
  • Q: What is the expected band size? Why is it different than the observed band size?
    A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:<br>1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
  • Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
    A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
  • Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
    A: Check our protocols under the tech support tab.
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