Overview

  • Product name
    Anti-Aquaporin 5 antibody [EPR3747]
    See all Aquaporin 5 primary antibodies
  • Description
    Rabbit monoclonal [EPR3747] to Aquaporin 5
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Aquaporin 5 aa 250-350. The exact sequence is proprietary.
    (Peptide available as ab214008)

  • Positive control
    • IHC-P: Human colon, lung and stomach tissue. WB: Fetal lung lysate. SW480 cell lysate. Flow Cyt: SW480 cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
  • Purity
    Tissue culture supernatant
  • Clonality
    Monoclonal
  • Clone number
    EPR3747
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab92320 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
Flow Cyt Use at an assay dependent concentration.
WB 1/1000 - 1/10000. Predicted molecular weight: 28 kDa.Can be blocked with Aquaporin 5 peptide (ab214008).
IHC-P 1/100 - 1/250. Antigen retrieval is recommended before commencing with IHC staining protocol. The use of an HRP/AP polymerized secondary antibody is recommended as stronger signals have been found using these.
ICC/IF 1/100 - 1/250.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      Forms a water-specific channel. Implicated in the generation of saliva, tears, and pulmonary secretions. Required for TRPV4 activation by hypotonicity (PubMed:16571723). Together with TRPV4, controls regulatory volume decrease in salivary epithelial cells (PubMed:16571723).
    • Involvement in disease
      Keratoderma, palmoplantar, Bothnian type
    • Sequence similarities
      Belongs to the MIP/aquaporin (TC 1.A.8) family.
    • Domain
      Aquaporins contain two tandem repeats each containing three membrane-spanning domains and a pore-forming loop with the signature motif Asn-Pro-Ala (NPA).
    • Cellular localization
      Apical cell membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • AQP 5 antibody
      • AQP-5 antibody
      • AQP5 antibody
      • AQP5_HUMAN antibody
      • Aquaporin 5 antibody
      • Aquaporin-5 antibody
      see all

    Images

    • Comparison of non-diseased human lung (left panel) and Type I PPB (right panel).

      Immunostaining for the distal lung Type I pneumocyte marker Aquaporin 5 using ab92320. Aquaorin 5 was expressed in distal alveoli of non-diseased lung tissue and in Type I PPB associated epithelium.

      The human Type I Pleuropulmonary Blastoma tissue samples, formalin fixed and paraffin embedded, were obtained through the genetic studies tissue bank of the International PPB registry.

    • In vitro characterization of key marker proteins AQP5, ANO1, α-SMA, and CEACAM5 in cultures of the SG (sweat glands) by immunofluorescence staining.

      Panel b shown: Stained secretory coil (sc) and duct (du) sections of native human sweat glands were analyzed in axillary skin tissue (a,d,g,j) compared to in situ glands (dashed lines) with outgrowing cells in 2D cultures (b,e,h,k) and 3D models (c,f,i,l). Polarization and differentiation of spheroids could be shown with luminal ANO1 (f) and basolateral α-SMA staining (i). Outgrown SG cells showed decreased marker expressions in the 2D cultures. Nuclei stained with DAPI.

    • ab92320 showing positive staining in human colon tissue.

    • ab92320 showing positive staining in human stomach tissue.

    • ab92320 at 1/100 dilution, staining Aquaporin 5 in paraffin embedded human lung tissue.

    • ab92320 showing negative staining in human brain tissue.

    • All lanes : Anti-Aquaporin 5 antibody [EPR3747] (ab92320) at 1/1000 dilution

      Lane 1 : Fetal lung lysate
      Lane 2 : SW480 (Human colorectal adenocarcinoma cell line) cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labeled goat anti-rabbit IgG at 1/1000 dilution

      Predicted band size: 28 kDa

    • Flow cytometry analysis of SW480 (Human colorectal adenocarcinoma cell line) cells labeling with unpurified ab92320 at 1/200 dilution (1 µg/ml) (Red).

      Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. Goat anti rabbit IgG Alexa Fluor® 488(ab150077) at a 1/2000 dilution, was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as a isotype control. Cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

    References

    This product has been referenced in:
    • Hwang S  et al. Carbonic anhydrase 12 mutation modulates membrane stability and volume regulation of aquaporin 5. J Enzyme Inhib Med Chem 34:179-188 (2019). Read more (PubMed: 30451023) »
    • Esghaei M  et al. Evaluation of Anticancer Activity of Camellia Sinensis in the Caco-2 Colorectal Cancer Cell Line Asian Pac J Cancer Prev 19:1697-1701 (2018). Read more (PubMed: 29938468) »
    See all 15 Publications for this product

    Customer reviews and Q&As

    1-10 of 13 Abreviews or Q&A

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Dog Tissue sections (intervertebral disc)
    Antigen retrieval step
    None
    Permeabilization
    No
    Specification
    intervertebral disc
    Blocking step
    1% BSA in 75% TBS 25% serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 25% · Temperature: 18°C
    Fixative
    10% neutral buffered formalin

    Abcam user community

    Verified customer

    Submitted Jun 05 2017

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Lung)
    Gel Running Conditions
    Reduced Denaturing (12% Bis-Tris NuPAGE)
    Loading amount
    50 µg
    Specification
    Lung
    Blocking step
    Odyssey TBS Blocking Buffer (proprietary) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 23°C

    Pete Serbedzija

    Verified customer

    Submitted Apr 08 2016

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (AQP5-EGFP transfected cells / rat lung slices)
    Permeabilization
    Yes - 0,2 % Saponin
    Specification
    AQP5-EGFP transfected cells / rat lung slices
    Blocking step
    Serum as blocking agent for 5 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Oct 01 2015

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Zytomed Systems HRP Polymer Kit Blocking Solution as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 25°C
    Antigen retrieval step
    None
    Sample
    Human Tissue sections (Lung tissue)
    Specification
    Lung tissue
    Permeabilization
    No
    Fixative
    HOPE-Fixation

    Abcam user community

    Verified customer

    Submitted Mar 12 2015

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (A549)
    Specification
    A549
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.25% Triton X-100/PBS

    Abcam user community

    Verified customer

    Submitted Apr 05 2013

    Answer

    Thank you for reply.

    I am afraid I will not be able to offer a discount code for the Anti-Prosurfactant Protein B antibody (ab104813) as I am not sure if the antibody is likely to cross react with the chicken protein.

    I can provide a discount code for the ab92320 as the immunogen used to raise this antibody shares 91% homology with a chicken protein (SwissProt reference http://www.uniprot.org/uniprot/Q683N0) and would therefore be predicted to react. However I could not do this for ab78486 as the immunogen used is taken from the C-terminal region of the rat protein which shares quite low homology with the chicken protein.

    In order to raise the discount code for ab92320 I will need the customers email address.

    I look forward to receiving your reply.

    Read More

    Answer

    Thank you for contacting us and sorry for the delay in getting back to you.

    In regards to the Anti-Prosurfactant Protein B antibody (ab104813), I was only able to locate a chicken protein, with the SwissProt reference http://www.uniprot.org/uniprot/E1BSP1 which was identified through performing a Blast of the protein sequence. This chicken protein only shares 24% homology with the human protein from which the immunogen is derived. I would therefore not be very sure that the antibody ab104813 would be able to detect a protein from the chicken samples. It may of course be that there are sequences from the chicken which have not been added to the SwissPort database yet.

    I was not able to find the antibody S841-04 which the customer had used in IHC. If they could sent the datasheet of this antibody I may be able to suggest a more promising alternative.

    I am able to provide a discount code for the customer for ab92320, however in order to do this I will need their direct email address so we can match them to the Abreview they will submit for the product.

    They do not however need this code directly for ordering so please let them go ahead and order and I can issues the discount code once you have replied with the email address.

    I look forward to receiving your reply.

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry
    Sample
    Human Cell (primary alveolar epithelial cells)
    Specification
    primary alveolar epithelial cells
    Fixative
    HOPE
    Permeabilization
    No
    Blocking step
    Zytomed Systems HRP Kit Blocking solution as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Jul 31 2012

    Answer

    Thank you for your enquiry.

    I can confirm that ab92320 antibody is sold as whole antiserum / ascites / tissue culture supernatant. Unpurified antibodies, such as those sold as tissue culture supernatant will not have a concentration stated on the datasheet. Antibody concentration is usually determined by protein assay, and serum / ascites / tissue culture supernatant will contain a lot of other proteins, which means the antibody quantification would not be accurate.

    I can confirm that for tissue culture supernatant, concentration of antibody is known to very between 1 - 3 mg/ml

    I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

    Read More

    Question
    Answer

    Thank you for your inquiry.

    I can confirm that this antibody does not react with mouse and rat brain, heart, kidney and spleen samples as tested by our lab in WB.

    I hope this information is helpful and wish you good luck with your experiments.

    Read More

    1-10 of 13 Abreviews or Q&A

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