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Anti-Calpain 5 antibody - Aminoterminal end domain I (ab28280)

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Overview

Product name

Anti-Calpain 5 antibody - Aminoterminal end domain I
See all Calpain 5 products (5) ...

Description

Rabbit polyclonal to Calpain 5 - Aminoterminal end domain I

Specificity

Does not cross react with the other calpain family members (Calpain 1, Calpain 2, Calpain 3, ncl 3, etc.).

Tested applications

WB, ICC/IFmore details

Cross reactivity

Reacts with

Mouse, Rat, Human

Immunogen

Synthetic peptide based on the N terminal end of the large subunit of Human Calpain 5. (Peptide available as ab41310.)

Epitope

Reacts to the first 30 amino acids of the "propeptide domain," (Domain-I) of Calpain 5, N terminal from the activation cleavage site.

General notes

Diluted antibody can be reused several times if not contaminated; store at 4°C with 0.05% Sodium Azide as preservative.

Properties

Form

Liquid

Storage instructions

Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

Storage buffer

Preservative: 0.05% Sodium Azide
Constituents: 50% Glycerol

Concentration

Concentration information loading...

Purity

Immunogen affinity purified

Purification notes

The antibody has been peptide-affinity purified.

Primary antibody notes

ab28280 does not recognize amino-processed Calpain 5, and can be used to discriminate between latent and amino-processed forms. ab28280 will also bind to the non-reduced protein, and thus may have application for immunoprecipitation, immunohistochemistry, and ELISA.

Clonality

Polyclonal

Isotype

IgG

Research areas

Cell Biology >> Proteolysis / Ubiquitin >> Proteolytic enzymes >> Cysteine protease >> Calpains
Signal Transduction >> Signaling Pathway >> Calcium Signaling >> Calpain

  • Immunocytochemistry/ Immunofluorescence-Calpain 5 antibody - Aminoterminal end domain I(ab28280)Immunocytochemistry/ Immunofluorescence-Calpain 5 antibody - Aminoterminal end domain I(ab28280) image (enlarge)

Applications

Show applications key

Our Abpromise guarantee covers the use of ab28280 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application notes

ICC/IF: Use at a concentration of 1 - 5 µg/ml.
WB: 1/1000 (colorimetric) - 1/5000 (chemiluminescent). Detects bands of approximately 73 and 68 kDa and a series of further cleaved forms when used against the reduced protein. Dilution optimised using Chromogenic detection.

Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.

Target

Relevance

Calpain proteinases are defined currently as papain like neutral proteases that are calcium activated. Calpains 1 and 2 are composed of a large subunit, which is proteolytically active, and a small subunit (also called calpain 4), or enhancer protein, that is not proteolytically active. The other calpain proteins identified to date are not known to require a small subunit. Domains in the large subunit include the amino terminal domain I, the proteinase domain II, domain III, and the EF hand domain IV (Domain T in calpains 5 & 6). Calpain 5, also known as the human orthologue of TRA3, a C. elegans protein involved in sex determination, is located on chromosome 11 in humans, near calpain 1. Calpain 5 is an intracellular, calcium dependent cysteine protease. Calpain 5 domain T does not have the standard calpain EF hand domain for binding calcium, making it unclear whether it is calcium regulated. It is also unclear if calpain 5 plays any role in mammalian sex determination, since the enzyme is found in colon, kidney, liver and trachea and other tissues, as well as testis.

Cellular localization

Cytoplasmic

Alternative names

  • CAPN 5 antibody
  • CAPN5 antibody
  • FLJ46245 antibody
  • HTRA3 antibody
  • nCL3 antibody
  • New calpain 3 antibody
see all

Database links

see all

Anti-Calpain 5 antibody - Aminoterminal end domain I images:

  Immunocytochemistry/ Immunofluorescence-Calpain 5 antibody - Aminoterminal end domain I(ab28280)

Immunocytochemistry/ Immunofluorescence-Calpain 5 antibody - Aminoterminal end domain I(ab28280)

ICC/IF image of ab28280 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab28280, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Protocols

  • Recommended WB protocol

    • References for Anti-Calpain 5 antibody - Aminoterminal end domain I (ab28280)

    ab28280 has not yet been referenced specifically in any publications.

    Publishing research using ab28280? Please let us know so that we can cite the reference in this datasheet

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"