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).
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.
The C28R2 sequence is very conserved in various isoforms among various species.
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：human
Grade：Serum
Purity：Serum
Form：Supplied as a liquid, neat serum, 40% glycerol.
Specificity：The 28 AA human C28R2 sequence is 100% conserved in human Ca+-ATPase isoforms 1, 2, 3b, 96% in 1b, 92% in 4 and 85% in 3a/3b. It is also 100% conserved in mouse and rat isoform 2. A high degree of conservation (77-100%) is also seen in Ca+-ATPase isoforms from many other species. Antibody crossreactivity of C28R2 or its exchange inhibitory properties in various species has not been studied. Control peptides, because of their low MW (<3kD), are not suitable for Western. It should be used for ELISA or antibody blocking to confirm antibody specificity
Isotype：IgG
Calc Applications Abbrev：E WB
Calc Crossreactivity：Hu Mo Rt
Immunogen：The 28 AA Peptide sequence (~CT, cytoplasmic) corresponding to human Ca+-ATPases, autoinhibitory domain (1). The peptide was coupled to KLH
