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K-Cl Cotransporters (KCC1-4)
Antibodies
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. Since cytoplasmic
electroneutrality is maintained under normal physiological environment,
changes in cellular chloride level is accompanied by total cell solute
content. Because of high water permeability of cell membranes, changes in
cell solute content are accompanied by changes in cellular volume. In order
to maintain constant cell volume and prevent swelling, cells must expend
energy through primary and secondary active transport mechanisms. The cation
chloride cotransporters (CCC) protein family is invovled in the
electroneutral movement of ions across the cell membrane.
Three groups of proteins have been identified in
CCC
family based upon their structures, ligands, and inhibitors. These are the
thiazide-sensitive Na+-Cl-cotransporters (NCC or TSC), the loop
diuretics-sensitive Na+-K+-Cl- (NKCC) cotransporters (NKCC1/BSC2 and
NKCC2/BSC1), and the
K+-Cl--cotransporters (KCC).
The K-Cl cotransporters mediate the coupled movement of K+ and Cl- ions
across the cell membranes. This transport process is involved in the
regulatory volume decrease in response to cell swelling in red cells and
vectorial movement of Cl- ions across the kidney epithelia. The net
direction of this transport is out of the cells. At least four isoforms of
KCC identified (KCC1, KCC2, KCC3,
and KCC4) have been identified and functionally
characterized. KCC are predicted to have the same protein structure: 12
transmembrane domains with a large extracellular loop with potential
N-glycosylation sites, and the cytoplasmic N and C-termini.
KCC1
(human/rat/mouse 1085 aa) is expressed in both erythroid and non-erythroid
cells and tissues. It is activated by cell swelling. KCC2 (rat 1116 aa) is
not activated by swelling and its expression is restricted to brain.
KCC2 may
function in buffering of external K+ within the central nervous system, in
addition to maintain the transmembrane Cl- gradient.
KCC3 is ~77%
identical to KCC1. Alternative splicing of KCC3 transcripts generates KCC3
with different N-termini (1099 aa and 1150 aa). It is highly expressed in
brain, heart, skeletal muscle, and kidney. KCC3 is modulated by VEGF and
TNF-alpha. Most recently, KCC4
(mouse 1083 aa, human 1083 aa) have been cloned that are 65-71% identical to
KCC1/KCC2. It is highly expressed in heart and kidney.
ADI has produced highly specific
rabbit antibodies to KCC1-4
using peptide sequences specific to each KCC. These
antibodies should be useful in studying the physiological roles of various
KCCs.
|
Items |
Antigen peptide location |
Antibody
Host |
Ab
Crossreactivity |
Neat
Antisera Cat #
(100 ul) |
Aff.
Pure Ab
Cat #
(100 ug) |
* Control
Peptide Cat#
(100 ug) |
|
rKCC1 |
r, 21 aa ~NT |
Rb |
m, r, h, rb |
KCC11-S |
KCC11-A |
KCC11-P |
|
rKCC2 |
r, 18 aa ~NT |
Rb |
r, m (h?) |
KCC21-S |
KCC21-A |
KCC21-P |
|
hKCC3 (Ab#1) |
h, 19 aa ~NT |
Rb |
h (m, r?) |
KCC31-S |
KCC31-A |
KCC31-P |
|
hKCC3a (Ab#2) |
h, 17 aa ~CT |
Rb |
h (m, r?) |
KCC32-S |
KCC32-A |
KCC32-P |
|
mKCC4 |
m, 16 aa ~NT |
Rb |
h, m |
KCC41-S |
KCC41-A |
KCC41-P |
|
Control Rabbit
IgG (non-immmune) |
For use in
ELISA, Western, Immunohisto. |
20009-1 (1
mg) |
m=mouse; r=rat; h=human;
c=chicken; f=frog; ~CT or ~NT=near C or N-terminus.
"Neat Antisera" are the
unpurified antiserum and it is suitable for ELISA and Western.
"Affinity pure"
antibodies have been over the antigen-affinity column
and recommended for immunohistochemical applications.
"Control peptides" can not
be used for Western as they are very short peptides. They are
intended for ELISA or antibody competition studies.
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