Anti-ATP6V1B2 antibody (ab73404)
Key features and details
- Rabbit polyclonal to ATP6V1B2
- Suitable for: IP, IHC-P, WB, ICC/IF
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-ATP6V1B2 antibody
See all ATP6V1B2 primary antibodies -
Description
Rabbit polyclonal to ATP6V1B2 -
Host species
Rabbit -
Tested applications
Suitable for: IP, IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Cow, Cynomolgus monkey, Orangutan
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 450 to the C-terminus of Mouse ATP6V1B2.
Read Abcam's proprietary immunogen policy (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.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
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 (1) 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.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
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Function
Non-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 similarities
Belongs to the ATPase alpha/beta chains family. -
Cellular localization
Endomembrane system. Melanosome. Endomembrane. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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Database links
- Entrez Gene: 338082 Cow
- Entrez Gene: 526 Human
- Entrez Gene: 11966 Mouse
- Entrez Gene: 100173673 Orangutan
- Entrez Gene: 117596 Rat
- Omim: 606939 Human
- SwissProt: P31408 Cow
- SwissProt: P21281 Human
see all -
Alternative names
- ATP6B1B2 antibody
- ATP6B2 antibody
- ATP6V1 B2 antibody
see all
Images
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Western blot - Anti-ATP6V1B2 antibody (ab73404)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.
Secondary
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. -
- Anti-ATP6V1B2 antibody (ab73404)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 -
Immunocytochemistry/ Immunofluorescence - Anti-ATP6V1B2 antibody (ab73404)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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V1B2 antibody (ab73404)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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V1B2 antibody (ab73404)This image is courtesy of an anonymous Abreviewab73404 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.
Protocols
Datasheets and documents
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Datasheet download
References (15)
ab73404 has been referenced in 15 publications.
- Evans HT et al. Cell-specific non-canonical amino acid labelling identifies changes in the de novo proteome during memory formation. Elife 9:N/A (2020). PubMed: 31904341
- Zhu M et al. Extracellular Glutamate-Induced mTORC1 Activation via the IR/IRS/PI3K/Akt Pathway Enhances the Expansion of Porcine Intestinal Stem Cells. J Agric Food Chem 67:9510-9521 (2019). PubMed: 31382738
- Zhao W et al. A subunit of V-ATPases, ATP6V1B2, underlies the pathology of intellectual disability. EBioMedicine 45:408-421 (2019). PubMed: 31257146
- A M et al. A complex containing lysine-acetylated actin inhibits the formin INF2. Nat Cell Biol 21:592-602 (2019). PubMed: 30962575
- Li M et al. Transient Receptor Potential V Channels Are Essential for Glucose Sensing by Aldolase and AMPK. Cell Metab 30:508-524.e12 (2019). PubMed: 31204282
Images
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Western blot - Anti-ATP6V1B2 antibody (ab73404)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.
Secondary
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. -
- Anti-ATP6V1B2 antibody (ab73404)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 -
Immunocytochemistry/ Immunofluorescence - Anti-ATP6V1B2 antibody (ab73404)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.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V1B2 antibody (ab73404)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.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V1B2 antibody (ab73404) This image is courtesy of an anonymous Abreviewab73404 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.
