Anti-ATP6V1A Antibody Picoband™

Product Name

Anti-ATP6V1A Antibody Picoband™

View all ATP6V1A Antibodies

SKU/Catalog Number

A10401-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-ATP6V1A Antibody Picoband™ catalog # A10401-1. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat Monkey.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-ATP6V1A Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A10401-1)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human ATP6V1A recombinant protein (Position: A37-D617).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A10401-1 is reactive to ATP6V1A in Human, Monkey, Mouse, Rat

Applications

A10401-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

70 kDa

Calculated molecular weight

68.304kDa

Background of ATP6V1A

V-type proton ATPase catalytic subunit A is an enzyme that in humans is encoded by the ATP6V1A gene. 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.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.25 μg/ml, Human, Mouse, Rat, Monkey
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of ATP6V1A using anti-ATP6V1A antibody (A10401-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human placenta tissue lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: monkey COS-7 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat kidney tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse kidney tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-ATP6V1A antigen affinity purified polyclonal antibody (Catalog # A10401-1) at 0.25 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ATP6V1A at approximately 70 kDa. The expected band size for ATP6V1A is at 70 kDa.

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Figure 2. IF analysis of ATP6V1A using anti-ATP6V1A antibody (A10401-1).
ATP6V1A was detected in an immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-ATP6V1A Antibody (A10401-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of U87 cells using anti-ATP6V1A antibody (A10401-1).
Overlay histogram showing U87 cells stained with A10401-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATP6V1A Antibody (A10401-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For ATP6V1A (Source: Uniprot.org, NCBI)

Alternative Names

70kD, isoform 1; ATP6A1; ATP6V1A1ATPase, H+ transporting, lysosomal, subunit A1; ATPase, H+ transporting, lysosomal (vacuolar proton pump), alpha polypeptide; ATPase, H+ transporting, lysosomal 70kDa, V1 subunit A; EC 3.6.3; EC 3.6.3.14; H(+)-transporting two-sector ATPase, subunit A; H+-transporting ATPase chain A, vacuolar (VA68 type); HO68; VA68; vacuolar ATP synthase catalytic subunit A, ubiquitous isoform; Vacuolar ATPase isoform VA68; vacuolar proton pump alpha subunit 1; Vacuolar proton pump subunit alpha; V-ATPase 69 kDa subunit 1; V-ATPase 69 kDa subunit; V-ATPase A subunit 1; V-ATPas ATP6V1A ARCL2D, ATP6A11, DEE93, HO68, IECEE3, VA68, VPP2, Vma1, ATP6V1A ATPase H+ transporting V1 subunit A V-type proton ATPase catalytic subunit A|ATPase, H+ transporting, lysosomal 70kDa, V1 subunit A|ATPase, H+ transporting, lysosomal, subunit A1|H(+)-transporting two-sector ATPase, subunit A|H+-transporting ATPase chain A, vacuolar (VA68 type)|V-ATPase 69 kDa subunit 1|V-ATPase A subunit 1|V-ATPase subunit A|vacuolar proton pump alpha subunit 1|vacuolar proton pump subunit alpha

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