Technical Data
B0094-50B
Barttin
Description:
Chloride is a critical component of all living cells. Voltage-gated chloride channels regulate cellular traffic of chloride ion. The chloride channels (CIC or CLC) performs several functions including the regulation of cell volume, membrane potential stabilization, signal transduction, and transepithelial transport. Mutations in CIC genes have been linked with several human diseases including myotonias (Thomsen's disease), cystic fibrosis, Bartters syndrome type III, Dent's disease, and X-linked recessive nephrolithiasis. In mammals, CLC proteins form a superfamily of at least 9 different genes (CLC1-7 also known as CLCN1-7 and CLK1-2 or CLCKa and CLCKb). Additional forms of these proteins are obtained by alternative splicing. All CLC proteins (~700-1000 aa) are predicted to contain 10 (possibly 12) transmembrane domains. Except CLC-1 and CLC-K1/K2 that are specific for kidney, most other CLC are widely distributed in various tissues.

Bartter syndrome is an autosomal recessive disorder defined by hyokalemic metabolic alkalosis, elevated plasma renin activity and hyperaldosteronism with normal blood pressure and altered prostaglandin metabolism. Mutations in the NKCC2, K+-channel ROMK, and CLCNKB genes are associated with Bartter syndrome type1-3, respectively. BSND encodes a protein known as barttin, which contains 2 TM and is expressed in thin limb and thick ascending limb of the loop of Henle in the kidney, and in the dark cells of the inner ear. BSND gene is mutated in Bartter syndrome with sensorineural deafness. Barttin (mouse 307-aa, rat 308-aa, human 320-aa, chromosome 1p31-p32, ~40-42kD) acts as an essential b-subunit for CLC-Ka and CLC-Kb with which it co-localizes in basolateral membrane of renal tubules and of K-secreting epithelia of the inner ear.

Applications:
Suitable for use in ELISA, Western Blotting. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000-1:5000 for neat serum and 1-10ug/ml for affinity pure antibody using ECL.
ELISA: 1:10,000-50,000 for neat serum and 0.5-1ug/ml for affinity pure. Control peptide can be used to coat ELISA plates at 1ug/ml.

Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
100ul-20CBlue IceHumanRabbit
Concentration:
As reported
Immunogen:
A 16-aa Peptide sequence after the 2nd TM domain and in the cytoplasmic region of the human/mouse/rat Barttin (1) was synthesized, coupled to KLH
Purity:
Purified by immunoaffinity chromatography.
Form
Supplied as a liquid in PBS, pH 7.4 and 0.1% sodium azide, 40% glycerol.
Specificity:
The control peptide 100% conserved in mouse, rat and human CLC-1. No significant sequence homology is detected with other CLCs or other proteins. Because of the low MW (<3kD) of the control peptide, it is not recommended for Western. Control peptide, because of its low MW (<3kD), is not suitable for Western. It should be used for ELISA or antibody blocking experiments (use 5-10 ug control peptide per 1 ug of aff pure IgG or 1 ul antiserum) to confirm antibody specificity
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
(1) Birkenhager R et al (2001) Nat. genet. 29, 310-324; Estevez R et al (2001) Nature 414, 558-561; Brennan TM et al (1998) Am. J. Hum. Genet. 62, 355-361