Recombinant Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)
- Datasheet
- References (2)
- Protocols
Overview
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Product name
Anti-Calpain small subunit 1 antibody [EPR3323]
See all Calpain small subunit 1 primary antibodies -
Description
Rabbit monoclonal [EPR3323] to Calpain small subunit 1 -
Host species
Rabbit -
Specificity
Detects Calpain small subunit 1. -
Tested applications
Suitable for: WB, IHC-P, ICC, Flow Cytmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human Calpain small subunit 1 aa 50-150. The exact sequence is proprietary.
Database link: P04632 -
Positive control
- T47-D, 293T, A431 and Human spleen lysates. Human kidney tissue.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR3323 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab92356 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | 1/1000 - 1/2000. Predicted molecular weight: 28 kDa. | |
| IHC-P | 1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. The use of an HRP/AP polymerized antibody is recommended as these have been shown to give superior staining. |
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| ICC | 1/100 - 1/250. | |
| Flow Cyt | 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Function
Regulatory subunit of the calcium-regulated non-lysosomal thiol-protease which catalyzes limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction. -
Sequence similarities
Contains 5 EF-hand domains. -
Domain
The contact of the 5th EF-hand domain from each monomer allows the formation of the homodimer and also appears to mediate the contact between the large catalytic subunit and small regulatory subunit for the formation of the heterodimer.
EF-hand domains are paired. EF-hand 1 is paired with EF-hand 2 and EF-hand 3 is paired with EF-hand 4. The fifth EF-hand domain, left unpaired, does not bind the calcium but is responsible of the dimerization by EF-embrace. The first four EF-hand domains bind calcium, however it is not sure if the binding of EF-hand 4 to calcium is physiologically relevant. -
Cellular localization
Cytoplasm. Cell membrane. Translocates to the plasma membrane upon calcium binding. - Information by UniProt
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Database links
- Entrez Gene: 826 Human
- Entrez Gene: 12336 Mouse
- Entrez Gene: 29156 Rat
- Omim: 114170 Human
- SwissProt: P04632 Human
- SwissProt: O88456 Mouse
- SwissProt: Q64537 Rat
- Unigene: 515371 Human
see all -
Alternative names
- 30K antibody
- Calcium activated neutral proteinase antibody
- Calcium activated neutral proteinase small subunit antibody
see all
Images
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![Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)](https://www.abcam.com/ps/products/92/ab92356/Images/ab92356-317908-anti-calpain-small-subunit-1-antibody-epr3323-western-blot.jpg)
Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)All lanes : Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : CAPNS1 (Calpain small subunit 1) knockout HAP1 whole cell lysate
Lane 3 : HepG2 whole cell lysate
Lane 4 : HeLa whole cell lysate
Lysates/proteins at 40 µg per lane.
Predicted band size: 28 kDaLanes 1 - 4: Merged signal (red and green). Green - ab92356 observed at 82 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab92356 was shown to specifically react with Calpain 1 in wild-type HAP1 cells as signal was lost in CAPNS1 (Calpain small subunit 1) knockout cells. Wild-type and CAPNS1 (Calpain small subunit 1) knockout samples were subjected to SDS-PAGE. ab92356 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)](https://www.abcam.com/ps/products/92/ab92356/Images/ab92356-107338-anti-calpain-small-subunit-1-antibody-epr3323-western-blot.jpg)
Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)All lanes : Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356) at 1/1000 dilution
Lane 1 : T47-D cell lysate
Lane 2 : 293T cell lysate
Lane 3 : A431 cell lysate
Lane 4 : Human spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit antibody at 1/2000 dilution
Predicted band size: 28 kDa -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)](https://www.abcam.com/ps/products/92/ab92356/Images/ab92356-107337-anti-calpain-small-subunit-1-antibody-epr3323-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)ab92356 at 1/100 dilution staining Calpain small subunit 1 in paraffin-embedded Human kidney
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Protocols
Datasheets and documents
References
This product has been referenced in:
- Seymen CM et al. Melatonin Modulates NMDA-Receptor 2B/Calpain-1/Caspase-12 Pathways in Rat Brain After Long Time Exposure to GSM Radiation. Turk Neurosurg N/A:N/A (2019). WB, IHC-P ; Rat . Read more (PubMed: 31608966) »
- Kim DH et al. Calpain 9 as a therapeutic target in TGFß-induced mesenchymal transition and fibrosis. Sci Transl Med 11:N/A (2019). Read more (PubMed: 31316008) »
