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

1-10 of 11 Q&A

Question
Answer

We recently compared this antibody against two other antibodies to Aquaporin 4 and they all gave a single band at about 46kDa. However a doublet at 46/48kDa is commonly seen when blotting for this target so if this is what you are seeing then I feel this data to be reasonable.

Read More

Answer

Thank you for your further questions.

I can confirm the following prices:
CD14 antibody [5A3B11B5] (ab36595) 100 ul £227.00
CD14 antibody [7] (ab49755) 200 ug £227.00
Delivery: £16.00 to United Kingdom

This information can be found on the individual online datasheets. Please ensure toselect the correct countryfrom thedropdown menu at the top right corner of the webpage. The correct price for the product and delivery charge in the UK will then show at the top of the page.

Regarding which of these antibodies is best for parafin embedded eye sections, I am sorry that neither of these CD14 antibodies have specifically been tested on eye sections as far as we are aware. I would like to reassure you that our antibodies have all been tested successfully and would be covered by our 6 month guarantee in the applications and specieslisted on the datasheets. Therefore, any CD14 antibody tested in thespecies and application you are using should be suitable for your applications. Also, although we produce many in-house antibodies, we obtain other antibodies from a wide range of sources. Therefore, comparison experiments will not often have been done.

If you wish to be more certain, I can suggest choosing the antibody that has been best characterized. By this I mean the antibody that has been tested in the most applications and has the most data available (on the datasheet).

I hope this will be helpful. Should you have any further questions please do not hesitate to ask.

Read More

Answer

Thank you for your enquiry which has been forwarded to the scientific support team.

I believe our customer service team have already provided a quotation for you. I can confirm that prices are also available on the online datasheets.

Regarding the IHC application, I can confirm thatab46182is not tested in paraffin embedded sections, this would be indicated by 'IHC-P'.

I am pleased to let you know that Epitomics is now an Abcam company, and ab128906 5582-1 Anti-Aquaporin 4 antibody [EPR7040] is tested in IHC-P and in rat and human samples. so this is suitable and covered by our 6 month guarantee for IHC-P.

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

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

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

Question
Answer

Thank you for contacting us.

The bands could be the isoforms. Please check the Swissprot database http://www.uniprot.org/uniprot/P55087.

We also recommend checking the publications and reviewing other authors results.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for contacting us.

We realize that our products can be considered to have premium price point. However, we feel that the product and service which we offer are of the highest possible quality. Under our Abpromise guarantee, ourproducts are guaranteed to work in the manner stated on the datasheetfor a period of six months. If you experience any difficulties using our product and if you have purchased this product within the last 180 days, we are happy to provide scientific support, refund or replacement.

We do offer bulk discounts for orders of 10 units or more, and often run promotions for a number of product types. Please see our website or speak with customer service representative for more information about these.

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Read More

Answer

Thank you for contacting us. I am glad to hear that this replacement product is performing better than the last product.

Many factors can alter the predicted molecular weight of a protein. These may include post-translational modifications, protein processing, isoforms from alternative splicing, multimer formation, and amino acid charge.

Non-specific binding of an antibody to proteins other than the antigen can sometimes occur. This is usually more common with polyclonal antibodies, but can also occur with monoclonals as well.

An immunizing peptide blocking experiment can be performed to determine which band or staining is specific. As you have mentioned, this product's immunogen is available for use as a blocking peptide. I would recommend using this if you are concerned about the specificity of this product.

In case you would like more information regarding the use of blocking peptides, I have included Abcam's protocol for blocking with the immunizing peptide.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your enquiry.

I would like to reassure you that our antibodies have all been tested successfully and would be covered by our guarantee in the applications listed on the datasheets. Therefore, any Aquaporin 4 antibody tested in the species and application you are using should be suitable for your applications.

Also, although we produce many in-house antibodies, we obtain other antibodies from a wide range of sources. Therefore, comparison experiments will not often have been done. I would recommend that when a user is deciding upon a product that they look for those products which have Abreviews and have been used in publications, if possible. When choosing an antibody to be used in Western blot or Flow Cytometry I would also recommend choosing a product which has either a peptide available or the peptide sequence published as the peptide can be used for blocking if necessary.

Please let me know which product your customer would prefer. I look forward to hearing from you. Should you have any further questions please do not hesitate to ask.

Thank you for contacting us.






I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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1-10 of 11 Q&A

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