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Mouse Anti-Calpain 2 antibody
Mouse Anti-Calpain 2 antibody
Calcium activated neutral proteinase 2; Calcium activated neutral proteinase; CALP80; Calpain 2 (m/II) large subunit; Calpain 2 catalytic subunit; Calpain 2 large catalytic subunit; Calpain 2 large subunit; Calpain large polypeptide L2; Calpain M type; Ca
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Details

Product Name Calpain 2
Chinese Name 钙激活的中性蛋白酶2单克隆抗体
Alias Calcium activated neutral proteinase 2; Calcium activated neutral proteinase; CALP80; Calpain 2 (m/II) large subunit; Calpain 2 catalytic subunit; Calpain 2 large catalytic subunit; Calpain 2 large subunit; Calpain large polypeptide L2; Calpain M type; Calcium-activated neutral proteinase 2; CALP80; Calpain 2, large [catalytic] subunit; Calpain M-type; Calpain-2 catalytic subunit; Calpain-2 large subunit; CAN2_MOUSE; M-calpain; Millimolar-calpain; Calpain2; CANP 2; CANP L2; CANP2; CANPL 2; CANPL2; CANPml; Capa2; CAPN 2; CAPN2; FLJ39928; M calpain; M calpin; M type; mCANP; Millimolar calpain.  
Research Area Tumour  Cell biology  Signal transduction  The new supersedes the old  
Immunogen Species Mouse
Clonality Monoclonal
Clone NO. D2S3
React Species Human, 
Applications WB=1:500-1000 IHC-P=1:25 Flow-Cyt=1:100 (Paraffin sections need antigen repair)
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
Theoretical molecular weight 78kDa
Cellular localization cytoplasmic The cell membrane 
Form Liquid
Concentration 1mg/ml
immunogen KLH conjugated synthetic peptide derived from human Calpain 2 
Lsotype IgG2b
Purification affinity purified by Protein G
Buffer Solution 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
Storage Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
Attention This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
PubMed PubMed
Product Detail Calpain, and m calpain, also known as Calpain 2, are intracellular, calcium dependent cysteine proteases. Mu calpain has a micromolar sensitivity (thus the mu) as compared to the millimolar calcium sensitivity of m calpain. Both Calpains 1 and 2 are composed of an 80 kD subunit and a 30 kD subunit. Whereas the 30 kDa subunit is shared by both enzymes, the larger catalytic subunits are different and exhibit the distinct Ca++ requirements that are suggested by their names.
The calpains have papain like activity, thus the pain nomenclature. Both Calpain 1 and Calpain 2 are ubiquitously expressed, and are countered by the endogenous calpain inhibitor, calpastatin.
Other calpain family members (calpain 94, ncl2, ncl3, etc) have more limited tissue distribution, and perhaps different functions. The calpain family members consist of a common small subunit (Calpain 4), and a large variable subunit. It is not clear that all calpains contain 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. The calpains appear to serve multiple physiological roles, and ideas concerning the functions of these enzymes are in a state of rapid flux.

Function:
Calcium-regulated non-lysosomal thiol-protease which catalyze limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction.

Subunit:
Forms a heterodimer with a small (regulatory) subunit (CAPNS1).

Subcellular Location:
Cytoplasm. Cell membrane. Note=Translocates to the plasma membrane upon Ca(2+) binding.

Tissue Specificity:
Ubiquitous.

Similarity:
Belongs to the peptidase C2 family.
Contains 1 calpain catalytic domain.
Contains 2 EF-hand domains.

SWISS:
P17655

Gene ID:
824

Database links:

Entrez Gene: 824 Human

SwissProt: P17655 Human



Product Picture
Sample:
Lane 1: A549 cell lysates
Lane 2: A431 cell lysates
Lane 3: SK-BR-3 cell lysates
Primary: Anti-Calpain 2 (SLM-51592M) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 78 kD
Observed band size: 78 kD
Paraformaldehyde-fixed, paraffin embedded (Human stomach section); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Calpain 2) Monoclonal Antibody, Unconjugated (SLM-51592M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Blank control: U87MG.
Primary Antibody (green line): Mouse Anti-CAPN2 antibody (SLM-51592M)
Dilution: 1:100;
Isotype Control Antibody (blue line): Mouse IgG Secondary Antibody : Goat anti-mouse IgG-AF488
Dilution: 1:400 Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with PBST for 20 min at room temperature.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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