Overview

  • Product nameAnti-ATP6V1B2 antibody
    See all ATP6V1B2 primary antibodies
  • Description
    Rabbit polyclonal to ATP6V1B2
  • Tested applicationsSuitable for: IP, IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Cynomolgus Monkey, Orangutan
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 450 to the C-terminus of Mouse ATP6V1B2.

    (Peptide available as ab74898.)

  • Positive control
    • This antibody gave a positive signal in the following tissue lysates: Rat hippocampus; Mouse hippocampus; Mouse kidney; Mouse testis; Human kidney; Human testis.

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • 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
  • Database links
  • Alternative names
    • ATP6B1B2 antibody
    • ATP6B2 antibody
    • ATP6V1 B2 antibody
    • ATP6V1B 2 antibody
    • ATP6V1B2 antibody
    • ATPase H+ transporting lysosomal 56/58kDa V1 subunit B isoform 2 antibody
    • ATPase H+ transporting lysosomal 56/58kDa V1 subunit B2 antibody
    • ATPase H+ transporting, lysosomal (vacuolar proton pump) beta polypeptide 56/58kD isoform 2 antibody
    • ATPase, H+ transporting, lysosomal V1 subunit B2 antibody
    • brain isoform antibody
    • Endomembrane proton pump 58 kDa subunit antibody
    • H+ transporting two sector ATPase antibody
    • HO 57 antibody
    • HO57 antibody
    • V ATPase B2 subunit antibody
    • V ATPase subunit B 2 antibody
    • V type proton ATPase subunit B, brain isoform antibody
    • V-ATPase subunit B 2 antibody
    • V-type proton ATPase subunit B antibody
    • Vacuolar ATP synthase subunit B brain isoform antibody
    • Vacuolar H+ ATPase 56,000 subunit antibody
    • Vacuolar H+ ATPase 56000 subunit antibody
    • Vacuolar H+ATPase B2 antibody
    • Vacuolar proton pump subunit B 2 antibody
    • VAT B2 antibody
    • VATB 2 antibody
    • VATB antibody
    • VATB2 antibody
    • VATB2_HUMAN antibody
    • Vma 2 antibody
    • Vma2 antibody
    • VPP 3 antibody
    • VPP3 antibody
    see all

Anti-ATP6V1B2 antibody images

  • All lanes : Anti-ATP6V1B2 antibody (ab73404) at 1 µg/ml

    Lane 1 : Hippocampus (Mouse) Tissue Lysate
    Lane 2 : Kidney (Mouse) Tissue Lysate
    Lane 3 : Testis (Mouse) Tissue Lysate
    Lane 4 : Human testis tissue lysate - total protein (ab30257)
    Lane 5 : Human kidney tissue lysate - total protein (ab30203)
    Lane 6 : Rat Hippocampus Tissue Lysate
    Lane 7 : Hippocampus (Mouse) Tissue Lysate
    Lane 8 : Kidney (Mouse) Tissue Lysate
    Lane 9 : Testis (Mouse) Tissue Lysate
    Lane 10 : Human testis tissue lysate - total protein (ab30257)
    Lane 11 : Human kidney tissue lysate - total protein (ab30203)
    Lane 12 : Rat Hippocampus Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    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
  • 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.

    See Abreview

References for Anti-ATP6V1B2 antibody (ab73404)

This product has been referenced in:
  • Rebsamen M  et al. SLC38A9 is a component of the lysosomal amino acid sensing machinery that controls mTORC1. Nature 519:477-81 (2015). Read more (PubMed: 25561175) »
  • Arnandis T  et al. Calpains mediate epithelial-cell death during mammary gland involution: mitochondria and lysosomal destabilization. Cell Death Differ : (2012). Read more (PubMed: 22555453) »

See all 4 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (incisors)
Specification incisors
Fixative Formaldehyde
Antigen retrieval step None
Permeabilization Yes - 0.1% triton x-100
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Feb 04 2013

Thank you for contacting Abcam.

The exact immunogen sequence for ab73404, but I can say the antibody will bind between amino acids ***.

Since it is polyclonal antibody, it will most likely bind to multiply sites within that region. ...

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Thank you for contacting us.Generally polyclonal antibodies are not epitope tested because they will bind many different places and not at a single site like a monoclonal antibody will do.

I hope this information is helpful to you. Please do n...

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