In cardiac myocytes, Ca(2+) concentrations alternate between high levels during contraction and low levels during relaxation. The increase in Ca(2+) concentration during contraction is primarily due to release of Ca(2+) from intracellular stores. However, some Ca(2+) also enters the cell through the sarcolemma(plasma membrane). During relaxation, Ca(2+) is sequestered within the intracellular stores. To prevent overloading of intracellular stores, the Ca(2+) that entered across the sarcolemma must be extruded from the cell. The Na(+)-Ca(2+) exchanger is the primary mechanism by which the Ca(2+) is extruded from the cell during relaxation. In the heart, the exchanger may play a key role in digitalis action. The exchanger is the dominant mechanism in returning the cardiac myocyte to its resting state following excitation.[supplied by OMIM].
Subcellular Location:
Cell membrane.
Tissue Specificity:
Expressed in cardiac sarcolemma, brain, kidney, liver, pancreas, skeletal muscle, placenta and lung.
Similarity:
Belongs to the sodium/potassium/calcium exchanger family. SLC8 subfamily. Contains 2 Calx-beta domains.
SWISS:
P32418
Gene ID:
6546
Database links:
Entrez Gene: 6546 Human
Entrez Gene: 20541 Mouse
Entrez Gene: 100328570 Rabbit
Entrez Gene: 29715 Rat
Omim: 182305 Human
SwissProt: P23685 Dog
SwissProt: P48766 Guinea pig
SwissProt: P32418 Human
SwissProt: P70414 Mouse
SwissProt: Q01728 Rat
Unigene: 31961 Human
Unigene: 468274 Human
Unigene: 728911 Human
Unigene: 265834 Mouse
Unigene: 118972 Rat
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Sample:
Lane 1: U251 (Human) Cell Lysate at 30 ug
Lane 2: A431 (Human) Cell Lysate at 30 ug
Lane 3: A549 (Human) Cell Lysate at 30 ug
Primary: Anti-NCX1 (SL1550R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 120 kD
Observed band size: 120 kD
Sample:
HL-60(Human) Cell Lysate at 30 ug
K562(Human) Cell Lysate at 30 ug
Primary: Anti-NCX1 (SL1550R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 106 kD
Observed band size: 118 kD
Sample:
293T(Human) Cell Lysate at 30 ug
HepG2(Human) Cell Lysate at 30 ug
Panc-1(Human) Cell Lysate at 30 ug
Primary: Anti-NCX1 (SL1550R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 106 kD
Observed band size: 118 kD
Paraformaldehyde-fixed, paraffin embedded (rat brain tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (NCX1) Polyclonal Antibody, Unconjugated (SL1550R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
U-937 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 20% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with CXCL2 Antibody(SL1550R) at 1:500 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Blank control:U937.
Primary Antibody (green line): Rabbit Anti-NCX1 antibody (SL1550R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
Protocol
The cells were incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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