Product nameAnti-ATP6V1B2 antibody
See all ATP6V1B2 primary antibodies
DescriptionRabbit polyclonal to ATP6V1B2
Tested applicationsSuitable for: IP, IHC-P, WB, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Cow, Cynomolgus monkey, Orangutan
- This antibody gave a positive signal in the following tissue lysates: Rat hippocampus; Mouse hippocampus; Mouse kidney; Mouse testis; Human kidney; Human testis.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab73404 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use a concentration of 5 µg/ml.|
|IHC-P||Use a concentration of 0.1 - 0.5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 57 kDa (predicted molecular weight: 57 kDa).|
|ICC/IF||Use a concentration of 5 µg/ml.|
FunctionNon-catalytic subunit of the peripheral V1 complex of vacuolar ATPase. V-ATPase is responsible for acidifying a variety of intracellular compartments in eukaryotic cells.
Sequence similaritiesBelongs to the ATPase alpha/beta chains family.
Cellular localizationEndomembrane system. Melanosome. Endomembrane. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
- Information by UniProt
- ATP6B1B2 antibody
- ATP6B2 antibody
- ATP6V1 B2 antibody
All lanes : Anti-ATP6V1B2 antibody (ab73404) at 1 µg/ml
Lane 1 : Hippocampus (Mouse) Tissue Lysate
Lanes 2 & 8 : Kidney (Mouse) Tissue Lysate
Lanes 3 & 9 : Testis (Mouse) Tissue Lysate
Lanes 4 & 10 : Human testis tissue lysate - total protein (ab30257)
Lanes 5 & 11 : Human kidney tissue lysate - total protein (ab30203)
Lanes 6 & 12 : Rat Hippocampus Tissue Lysate
Lane 7 : Hippocampus (Mouse) Tissue Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 57 kDa
Observed band size: 57 kDa
Additional bands at: 37 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 30 seconds
Lanes 1-6 were blocked with 5% BSA, Lanes 7-12 were blocked with 3% milk. Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
ATP6V1B2 was immunoprecipitated using 0.5mg Mouse Testis tissue lysate, 5µg of Rabbit polyclonal to ATP6V1B2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Testis tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab73404.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 57kDa; ATP6V1B2
ICC/IF image of ab73404 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab73404, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) PC12 cells at 5µg/ml.
IHC image of ATP6V1B2 staining in human hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab73404, 0.5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
ab73404 staining RAt incisor tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2.5% serum for 30 minutes at 22°C. Samples were incubated with primary antibody (1/1000 in 1% BSA/ 0.5% Triton X-100 in PBS) for 16 hours at 4°C. An undiluted peroxidase-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.
This product has been referenced in:
- Fassio A et al. De novo mutations of the ATP6V1A gene cause developmental encephalopathy with epilepsy. Brain 141:1703-1718 (2018). Read more (PubMed: 29668857) »
- Rahman N et al. Soluble adenylyl cyclase is essential for proper lysosomal acidification. J Gen Physiol 148:325-39 (2016). Read more (PubMed: 27670898) »