钙蛋白酶1(CAPN1)活性蛋白

[ PROPERTIES ]

Source: Prokaryotic expression. Host: E. coli

Residues: Pro75~Asp356

Tags: N-terminal His-tag

Purity: >95%

Buffer Formulation: 20mM Tris, 150mM NaCl, pH8.0, containing 0.05% sarcosyl

and 5% trehalose. Applications: Cell culture; Activity Assays. (May be suitable for use in other assays to be determined by the end user.)

Predicted isoelectric point: 5.3

Predicted Molecular Mass: 33.5kDa

Accurate Molecular Mass: 33kDa as determined by SDS-PAGE reducing conditions.

[ USAGE ]

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0mg/mL. Do not vortex. 

[ STORAGE AND STABILITY ]

Storage: Avoid repeated freeze/thaw cycles. 

Store at 2-8^oC for one month. Aliquot and store at -80^oC for 12 months.

Stability Test: The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37^oC for 48h, and no obvious degradation and precipitation wereobserved. The loss rate is less than 5% within the expiration date underappropriate storage condition. 

[ SEQUENCE ]

[ ACTIVITY ] 

Calpain 1, Large Subunit (CAPN1) is an intracellular protease that requirescalcium for its catalytic activity. Calcium-regulated non-lysosomal thiol-proteasewhich catalyze limited proteolysis of substrates involved in cytoskeletal remodelingand signal transduction. It has broad endopeptidase specificity. Besides, SignalTransducer And Activator Of Transcription 3 (STAT3) has been identified as aninteractor of CAPN1, thus a binding ELISA assay was conducted to detect theinteraction of recombinant rat CAPN1 and recombinant rat STAT3. Briefly, CAPN1were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uLwere then transferred to STAT3-coated microtiter wells and incubated for 2h at37℃. Wells were washed with PBST and incubated for 1h with anti-CAPN1 pAb,then aspirated and washed 3 times. After incubation with HRP labelled secondaryantibody, wells were aspirated and washed 3 times. With the addition of substratesolution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stopsolution to the wells and read at 450nm immediately. The binding activity of ofCAPN1 and STAT3 was shown in Figure 1, and this effect was in a dosedependent manner.

[ IDENTIFICATION ]

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