ATP6V1A Recombinant Rabbit Monoclonal Antibody [PSH21-14]
cat.: HA751786
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat, Monkey |
| Applications: | WB, IF-Cell, IHC-Fr, IHC-P, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH21-14 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*PBS (pH7.4). |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 68 kDa |
| Isotype: | IgG |
| Positive control: | HeLa cell lysate, A431 cell lysate, NIH/3T3 cell lysate, COS-1 cell lysate, Mouse kidney tissue lysate, Rat kidney tissue lysate, Mouse brain tissue lysate, Rat brain tissue lysate, HeLa, NIH/3T3, human kidney tissue, mouse kidney tissue, rat kidney tissue. |
| Subcellular location: | Cytoplasm, cytosol, Cytoplasmic vesicle, secretory vesicle, clathrin-coated vesicle membrane, Lysosome. |
| Recommended Dilutions:
WB IF-Cell IHC-Fr IHC-P FC IP |
1:20,000 1:500 1:500 1:8,000 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P38606 Human | P50516 Mouse Entrez Gene: 685232 Rat |
| 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 |
Images
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Fig1:
Western blot analysis of ATP6V1A on different lysates with Rabbit anti-ATP6V1A antibody (HA751786) at 1/20,000 dilution. Lane 1: HeLa cell lysate Lane 2: A431 cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: COS-1 cell lysate Lane 5: Mouse kidney tissue lysate Lane 6: Rat kidney tissue lysate Lane 7: Mouse brain tissue lysate Lane 8: Rat brain tissue lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 68 kDa Observed band size: 68 kDa Exposure time: 6 seconds; ECL: K1801; 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 (HA751786) at 1/20,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of HeLa cells labeling ATP6V1A with Rabbit anti-ATP6V1A antibody (HA751786) 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 (HA751786) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
Immunocytochemistry analysis of NIH/3T3 cells labeling ATP6V1A with Rabbit anti-ATP6V1A antibody (HA751786) 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 (HA751786) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Application: IHC-Fr Species: Mouse Site: kidney Sample: Frozen section Antibody concentration: 1/500 Antigen retrieval: Not required |
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Fig5:
Application: IHC-Fr Species: Rat Site: kidney Sample: Frozen section Antibody concentration: 1/500 Antigen retrieval: Not required |
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Fig6:
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-ATP6V1A antibody (HA751786) 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 (HA751786) 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. |
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Fig7:
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-ATP6V1A antibody (HA751786) 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 (HA751786) 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. |
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Fig8:
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-ATP6V1A antibody (HA751786) 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 (HA751786) 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. |
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Fig9:
Flow cytometric analysis of HeLa cells labeling ATP6V1A. Cells were fixed and permeabilized. Then stained with the primary antibody (HA751786, 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 iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig10:
ATP6V1A was immunoprecipitated from 0.2 mg 293T cell lysate with HA751786 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751786 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: HA751786 IP in 293T cell lysate Lane 3: Rabbit IgG instead of HA751786 in 293T cell lysate Blocking/Dilution buffer: primary antibody dilution (K1803) Exposure time: 10 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.