Buffer Formulation	PBS, pH7.4, containing 0.01% SKL, 1mM DTT, 5% Trehalose and Proclin300.
Traits	Freeze-dried powder
Purity	> 97%
Isoelectric Point	7.1
Applications	Cell culture; Activity Assays.

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.

 

Image


Figure. SDS-PAGE; Sample: Active recombinant CLCA1, Human.

 

Figure. Western Blot; Sample: Recombinant CLCA1, Human.

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