WB
Western blot analysis of ATP6V1A on different lysates with Rabbit anti-ATP6V1A antibody at 1/20,000 dilution. Lysates/proteins at 10 µg/Lane. Exposure time: 6 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/20,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-ATP6V1A antibody at 1/8,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/8,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-ATP6V1A antibody at 1/8,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/8,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-ATP6V1A antibody at 1/8,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/8,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of HeLa cells labeling ATP6V1A with Rabbit anti-ATP6V1A antibody at 1/500 dilution. Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ATP6V1A antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.IF-F
Application: IHC-Fr. Species: Mouse. Site: kidney. Sample: Frozen section. Antibody concentration: 1/500. Antigen retrieval: Not required.IF-F
Application: IHC-Fr. Species: Rat. Site: kidney. Sample: Frozen section. Antibody concentration: 1/500. Antigen retrieval: Not required.FC
Flow cytometric analysis of HeLa cells labeling ATP6V1A. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
ATP6V1A was immunoprecipitated from 0.2 mg 293T cell lysate with ATP6V1A Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ATP6V1A Recombinant Rabbit Monoclonal Antibody at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: 293T cell lysate (input). Lane 2: ATP6V1A Recombinant Rabbit Monoclonal Antibody IP in 293T cell lysate. Lane 3: Rabbit IgG instead of ATP6V1A Recombinant Rabbit Monoclonal Antibody in 293T cell lysate. Blocking/Dilution buffer: primary antibody dilution. Exposure time: 10 seconds.| Product Name | ATP6V1A Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIF-FIHCIPWB |
| WB:1:20000 IHC:1:8000 ICC/IF:1:500 FC:1:1000 IF-F:1:500 IP:1-2μg/sample |
|
| Species Reactivity | HumanMonkeyMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | ATP6V1A |
|---|---|
| Gene Synonyms | HO68 VA68 VPP2 Vma1 DEE93 ARCL2D ATP6A1 IECEE3 ATP6V1A1 |
| Gene Full Name | ATPase H+ transporting V1 subunit A |
| Gene Summary | This gene encodes a component of vacuolar ATPase (V-ATPase), a multisubunit enzyme that mediates acidification of eukaryotic intracellular organelles. V-ATPase dependent organelle acidification is necessary for such intracellular processes as protein sorting, zymogen activation, receptor-mediated endocytosis, and synaptic vesicle proton gradient generation. V-ATPase is composed of a cytosolic V1 domain and a transmembrane V0 domain. The V1 domain consists of three A and three B subunits, two G subunits plus the C, D, E, F, and H subunits. The V1 domain contains the ATP catalytic site. The V0 domain consists of five different subunits: a, c, c', c", and d. Additional isoforms of many of the V1 and V0 subunit proteins are encoded by multiple genes or alternatively spliced transcript variants. This encoded protein is one of two V1 domain A subunit isoforms and is found in all tissues. Transcript variants derived from alternative polyadenylation exist. [provided by RefSeq, Jul 2008] |
| Alternative Names | V-type proton ATPase catalytic subunit A EC:7.1.2.2 V-ATPase subunit A V-ATPase 69 kDa subunit Vacuolar ATPase isoform VA68 Vacuolar proton pump subunit alpha ATP6V1A ATP6A1 ATP6V1A1 VPP2 |
| Molecular Weight(MW) | 68 kDa |
| Cellular Localization | Cytoplasm, cytosol, Cytoplasmic vesicle, secretory vesicle, clathrin-coated vesicle membrane, Lysosome. |
WB
Western blot analysis of ATP6V1A on different lysates with Rabbit anti-ATP6V1A antibody at 1/20,000 dilution. Lysates/proteins at 10 µg/Lane. Exposure time: 6 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/20,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-ATP6V1A antibody at 1/8,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/8,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-ATP6V1A antibody at 1/8,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/8,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-ATP6V1A antibody at 1/8,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/8,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling ATP6V1A with Rabbit anti-ATP6V1A antibody at 1/500 dilution. Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ATP6V1A antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
IF-F
Application: IHC-Fr. Species: Mouse. Site: kidney. Sample: Frozen section. Antibody concentration: 1/500. Antigen retrieval: Not required.
IF-F
Application: IHC-Fr. Species: Rat. Site: kidney. Sample: Frozen section. Antibody concentration: 1/500. Antigen retrieval: Not required.
FC
Flow cytometric analysis of HeLa cells labeling ATP6V1A. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
ATP6V1A was immunoprecipitated from 0.2 mg 293T cell lysate with ATP6V1A Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ATP6V1A Recombinant Rabbit Monoclonal Antibody at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: 293T cell lysate (input). Lane 2: ATP6V1A Recombinant Rabbit Monoclonal Antibody IP in 293T cell lysate. Lane 3: Rabbit IgG instead of ATP6V1A Recombinant Rabbit Monoclonal Antibody in 293T cell lysate. Blocking/Dilution buffer: primary antibody dilution. Exposure time: 10 seconds.| Application Notes | WB:1:20000 IHC:1:8000 ICC/IF:1:500 FC:1:1000 IF-F:1:500 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage Buffer | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
