Ca2+ plays a critical role in intracellular signaling.
Intracellular Ca2+ levels are tightly controlled by continuos removal of Ca2+ via ATP-driven Ca2+ pump in the endoplasmic reticulum and plasma membrane, and Ca2+ transport system, the Na+/Ca2+ exchangers (NCX), in the plasma membrane.
NCX can move Ca2+ either into or out of cells, depending on the net Na+, Ca2+, and K+ gradient across the membrane.
In most cells, 3 Na+ are exchanged for 1 Ca2+.
In mammals, at least 5 distinct genes code for the exchangers: Three NCX (NCX1, NCX2, and NCX3), and two in the NCKX family (NCKX1 and NCKX2).
NCX share significant sequence homology (~70%), display 11 TM domains, a large central, intracellular hydrophilic regulatory loop between TM5 and 6, extracellular N-terminus and cytoplasmic C-terminus.
The N-terminal signal peptide is cleaved off from the mature exchanger protein.
NCX contains a highly basic region in the large hydrophilic, intracellular loop called XIP (Exchange inhibitory peptide; RRLLFYKYVYKRYRAGKQRG 20 aa), that inhibits Na-Ca+ exchange in cardiac sarcolemmal vesicles and in other cells.
Little or no sequence identity is found between the NCX and the Ca-pump.
However, XIP also inhibits the Ca pumps with more or less same efficiency as C28R2 peptide sequence (LRRGQILWFRGLNRIQTQIRVVKAFRSS, 28 aa) corresponding to the autoinhibitory domain of Ca-pump.
Ca+-pump is a Mg+-dependent enzyme that catalyzes the hydrolysis of ATP with the transport of Ca+.
Plasma membrane Ca+-ATPase (calcium pump) exists in several isoforms (human isofroms 1, 1084 aa; 1b, 1220 aa; from 2, 198/1243 aa; 3a/b 1173/1220 aa; 4, 1205 aa) and numerous sub-isoforms have been identified in various species.
Ca+-pump displays 10 TM domains, with cytoplasmic N and C-termini.
C28R2 is located in within the cytoplasmic, C-terminal domain of Ca+-ATPases.
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 purified.
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 4°C for short-term only.
For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%).
Freeze at -20°C.
Aliquots are stable for at least 12 months at -20°C.
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.
仕様
Size：100ul
Host：rabbit
Source Antibody：rat
Grade：Serum
Purity：Serum
Form：Supplied as a liquid, neat serum, 40% glycerol.
Specificity：The 20 AA human sequence is 75-100% conserved in mouse Ca+-ATPase isoforms 1, 1b, 2, 3a/b and 4 in mouse, rat and human. 85% in 3a/3b. The control peptide is available to confirm specificity of antibodies
Isotype：IgG
Calc Applications Abbrev：E WB
Calc Crossreactivity：Hu Mo Rt
Immunogen：A 20 AA Peptide sequence near the N-terminus of mature Rat NCKX2 (1). The peptide was coupled to KLH
