Overview

  • Product name
  • Description
    Rabbit polyclonal to Aquaporin 4
  • Host species
    Rabbit
  • Specificity
    ab46182 is not suitable for staining Aquaporin 4 in IHC using paraffin embedded sections but some customers have successfully used the antibody in frozen sections (see submitted Abreviews). Please contact Scientific Support if you need further help.
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-FrFl, ICCmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
    Predicted to work with: Sheep, Chicken, Cow
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human Aquaporin 4.

    Read Abcam's proprietary immunogen policy (Peptide available as ab46181.)

  • Positive control
    • This antibody gave a positive signal in the following Tissue Lysates: Brain (Human), Heart (Human) and Brain (Rat).

Properties

  • 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
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab46182 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 48 kDa (predicted molecular weight: 35 kDa).Can be blocked with Human Aquaporin 4 peptide (ab46181). Abcam recommends using milk as the blocking agent.
ICC/IF Use at an assay dependent concentration.
IHC-FrFl Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.

Target

  • Function
    Forms a water-specific channel. Osmoreceptor which regulates body water balance and mediates water flow within the central nervous system.
  • Tissue specificity
    Brain - muscle >> heart, kidney, lung, and trachea.
  • 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).
  • Post-translational
    modifications
    Phosphorylation by PKC at Ser-180 reduces conductance by 50%. Phosphorylation by PKG at Ser-111 in response to glutamats increases conductance by 40%.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • AQP 4 antibody
    • AQP-4 antibody
    • AQP4 antibody
    • AQP4_HUMAN antibody
    • Aquaporin type 4 antibody
    • Aquaporin-4 antibody
    • Aquaporin4 antibody
    • HMIWC 2 antibody
    • HMIWC2 antibody
    • Mercurial insensitive water channel antibody
    • Mercurial-insensitive water channel antibody
    • MGC22454 antibody
    • MIWC antibody
    • WCH 4 antibody
    • WCH4 antibody
    see all

Images

  • All lanes : Anti-Aquaporin 4 antibody (ab46182) at 1 µg/ml (blocked with 3% milk)

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Brain (Rat) Tissue Lysate
    Lane 3 : Human heart tissue lysate - total protein (ab29431)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 35 kDa
    Observed band size: 48 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 22 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 12 minutes
  • ab46182 staining Aquaporin 4 in mouse RAW 264.7 cells by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed with paraformaldehyde and blocked with 10% serum for 20 minutes at 24°C. Samples were incubated with primary antibody (1/100 in PBS) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • Immunocytocchemical analysis of 33B cells originating from Rat nervous tissue oligodendroglioma, labelling AQP4 with ab46182 diluted 1/100. Cells were PFA fixed. Hematoxyling used to counterstained. Negative control given were ab46182 was not included.

    See Abreview

  • Immunohistochemical analysis of Pig brain tissue labelling Aquaporin 4 with ab46182 at 1/100. Tissue fixed by perfussion, antigen retrieval performed by microwaving in citric acid buffer. DAB staining. 

    See Abreview

References

This product has been referenced in:
  • Zheng Y  et al. miR-29a ameliorates ischemic injury of astrocytes in vitro by targeting the water channel protein aquaporin 4. Oncol Rep 41:1707-1717 (2019). Read more (PubMed: 30628716) »
  • Clayton AM  et al. Postpartum increases in cerebral edema and inflammation in response to placental ischemia during pregnancy. Brain Behav Immun 70:376-389 (2018). WB ; Rat . Read more (PubMed: 29588233) »
See all 19 Publications for this product

Customer reviews and Q&As

11-20 of 30 Abreviews or Q&A

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Neuro2a)
Loading amount
6 µg
Specification
Neuro2a
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10% gel)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Sep 11 2012

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (33B)
Specification
33B
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Aug 01 2012

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (RAW cell line)
Specification
RAW cell line
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Aug 01 2012

Application
Western blot
Sample
Rat Cell lysate - whole cell (B50 cell line)
Loading amount
6 µg
Specification
B50 cell line
Gel Running Conditions
Non-reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jul 25 2012

Application
Immunocytochemistry
Sample
Rat Cultured Cells (33B)
Specification
33B
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Jul 06 2012

Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Neuro2a cell line)
Specification
Neuro2a cell line
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Jul 03 2012

Question
Answer

Vielen Dank für Ihren Anruf.

Das Labor hat mir nun bestätigt, dass dieser Antikörper soeben wieder in Lager gebracht werden konnte. Leider hat diese neue Charge jedoch die Kontrolle zur Immunhistochemie nicht bestanden und wir könnendieseneueChargedemfall nicht zur Immunhistochemie empfehlen. Die Kontrolle im Western blot war jedoch positiv und wir garantieren diesen Antikörper zur Verwendung im Western blot. Das Datenblatt wurde auch dementsprechend nun aktualisiert. https://www.abcam.com/index.html?datasheet=46182 (or use the following: https://www.abcam.com/index.html?datasheet=46182).

Ich hoffe, Sie brauchen diesenAntikörper im Western blot. Falls Sie den Antikörper in anderen Applikation benutzen wollten, kontaktieren Sie mich bitte zuerst.

Ich hoffe diese Informationen sind Ihnen hilfreich. Bitte zögern Sie nicht, uns wieder zu kontaktieren sollten Sie Fragen oder Bedenken haben.

Read More
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain - hippocampus)
Specification
Brain - hippocampus
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 24°C

Abcam user community

Verified customer

Submitted May 28 2012

Answer

Thank you for your reply.


I can certainly understand that this must be very frustrating for the customer. Unfortunately, since we haven't tested the antibody in this application it is difficult to know if the problem is with the antibody or if the antibody simply isn't suitable.


Based on IHC-P or -Fr data, the antibody can be used up to 1:2000. Allowing less antibody to penetrate the tissue over a longer period of time may allow for more specific staining.


I hope this additional advice is helpful!

Read More

Question
Answer

Thank you for contacting Abcam regarding ab46182.


I am sorry that you have been experiencing difficulties with this antibody in IHC. I was hoping you could clarify some details of your experiment so that I may assist you further.


Are these tissues frozen or paraffin embedded? Have you tried another fixation method? Would you be able to provide an image of your staining?


I would recommend testing multiple dilutions as 1:200 may be giving high background. I would also recommend increasing to 10% serum for blocking. Antibody should be diluted in PBS + 5% BSA. I would also suggest using tween instead of triton for these steps to maintain the integrity of the tissue.


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions or concerns or if these suggestions do not improve your results with this antibody.

Read More

11-20 of 30 Abreviews or Q&A

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