Cat. # NKCC21-P, Rat NKCC2 Control Peptide, SIZE: 100 ug
Cat. # NKCC21-S, Rabbit Anti-rat NKCC2 antiserum, SIZE: 100 ul
Cat. # NKCC21-A, Rabbit Anti-rat NKCC2 antiserum (affinity pure), SIZE: 100 ug

Chloride is a critical component of all living cells. It is also the single most dominant diffusible anion inside of most cells - the others are mostly impermeable organic anions. The cation chloride cotransporters (CCC) protein family is involved in the electroneutral movement of ions across the cell membrane. It includes the thiazide-sensitive Na+-Cl-cotransporters (NCC or TSC), the loop diuretics-sensitive Na+-K+-Cl- (NKCC) cotransporters (NKCC1/CCC1/BSC2 and NKCC2/CCC2/BSC1), and the K+-Cl--cotransporters (KCC1-4). These co-transporters share a common predicted membrane topology, with 12 TM domains (~500 aa), and long hydrophilic, intracellular N-and C-termini containing regulatory phosphorylation sites. NKCC transport Na, K, and Cl ions into and out of a wide variety of epithelial and non-epithelial cells. The transport process is characterized by electroneutrality (almost always with stoichiometry of 1Na:1K:2Cl) and inhibition by the loop diuretic bumetanide, benzametanide, and furosemide.

Unlike NKCC1, NKCC2 (human 1099 aa, rat/mouse 1095 aa; calculated mol size of ~121 kDa; actual size ~150 kDa; ~45% identity with NKCC1) is most strongly expressed in the kidney (inner and outer stripes of medulla and cortical thick ascending limb), and macula densa. NKCC2 plays a critical role in transcellular absorption of Na+-Cl- by the medullary and cortical TALH, and a secondary role in the paracellular transport of Na-Ca and Mg. Mutations in the NKCC2 gene result in Bartter's syndrome, an inherited disease characterized by hypokalemic metabolic alkalosis, hypercalciuria, salt wasting, and volume depletion.

Source of Antigen and Antibodies

Antigen: 15aa peptide of Rat NKCC2; Designated (NKCC21-P).
Location: ~C-terminus, Cytoplasmic
Ab Host/type: Rabbit, polyclonal
Ab Format: Aff pure IgG (cat #NKCC21-A)

Recommended Usage

Western Blotting (1:1K-5K for neat serum and 1-10 ug/ml for affinity pure using Chemiluminescence technique).

ELISA (1:10K-1:100K; using 50-100 ng of control peptide/well).

Histochemistry & Immunofluorescence: Not tested. We recommend the use of affinity purified antibody at 2-20 ug/ml.

Specificity & Cross-reactivity

The 15 AA rat NKCC21-P control peptide 100% conserved in mouse, 87% in human and rabbit NKCC2. No significant sequence homology is detected with NKCC1 or TSC. Antibody cross-reactivity in various species has not been studied. The NKCC21-P control peptide, because of its small size (<2 kDa), is not suitable for Western. It should be used in ELISA, dot blot or to neutralize antibody and confirm specificity of antibody.

References:

(1). Gamba G et al (1994) J Biol. Chem. 269, 17713; Simon DB et al (1996) Nature Genet. 13, 183; Mount DB et al (1995) J Am. Soc. Nephrol. 6, 347; Igarashi P et al (1995) Am. J. Physiol. 269, F405; Haas M et al (1987) Am. J. Physiol. 253, C243; Mount DB et al (1998) J. Exptl. Biol. 201, 2091 (review).