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Active Chloride Channel Accessory 1 (CLCA1)

CaCC1; CLCRG1; Chloride Channel,Calcium Activated,Family Member 1; Calcium-activated chloride channel protein 1

ACTIVITY TEST

CLCA1 (Calcium-activated chloride channel regulator 1) is a member of the calcium sensitive chloride conductance protein family. This protein is expressed as a precursor protein that is processed into two cell-surface-associated subunits. It has been reported that CLCA1 activates calcium-dependent chloride channel through the interaction with TMEM16A (anoctamin-1). Besides, there exits similarities between human and mouse TMEM16A in amino acid sequence with the identity of 89.66%. Thus, a functional ELISA assay was conducted to detect the association of recombinant human CLCA1 with recombinant mouse TMEM16A. Briefly, CLCA1 were diluted serially in PBS with 0.01%BSA (pH 7.4). Duplicate samples of 100uL were then transferred to TMEM16A-coated microtiter wells and incubated for 2h at 37°C. Wells were washed with PBST and incubated for 1h with anti-CLCA1 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37°C. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of CLCA1 with TMEM16A was shown in Figure 1 and this effect was in a dose dependent manner.

USAGE

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

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

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°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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INCREMENT SERVICES

Magazine Citations
Cell Communication and Signaling CLCA1 suppresses colorectal cancer aggressiveness via inhibition of the Wnt/beta-catenin signaling pathway pubmed:28974231
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