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

21-30 of 30 Abreviews or Q&A

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.

Read More

Question

LOT NUMBER GR54893-1
ORDER NUMBER *********
DESCRIPTION OF THE PROBLEM Non-specific staining Abnormal staining pattern (it stains cell nucleus, istead of cell membrane as usual)
SAMPLE Mouses
PRIMARY ANTIBODY Primary antibody: Anti-Aquaporin4 Ab (Ab 46182) Concentration or dilution: x3000 and x500 dilution Diluent buffer: 5% goat serum Incubation time: 2hr wash Buffer: 0.1M phosphate burffered saline (+0.5% triton X-100) Number of washes: 3 times
DETECTION METHOD - POSITIVE AND NEGATIVE CONTROLS USED - ANTIBODY STORAGE CONDITIONS -20℃ FIXATION OF SAMPLE If yes: 4% Paraform aldehyde Fixation time: 2 hrs Fixation temperature: 4 ˚c
ANTIGEN RETRIEVAL none PERMEABILIZATION STEP Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers? yes Permeabilizing agent and concentration: 30% Sucrose, 4 ˚c overnight
BLOCKING CONDITIONS Blocking agent (eg BSA, serum…): goat serum (+0.5% triton X-100) Concentration: 5% Blocking time: 1hr Blocking temperature: room temperature Endogenous peroxidases blocked? no Endogenous biotins blocked? no
SECONDARY ANTIBODY Secondary antibody: Alexa Fluor 488(green) and 568(red) goat anti-rabbit IgG(H+L) Species: goat Isotype: IgG Reacts against: rabbit Concentration or dilution: x1000 dilution Diluent buffer: 5% goat serum Incubation time: 1hr wash Buffer: 0.1M phosphate burffered saline (+0.5% triton X-100) Number of washes: 3 times
HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4
HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

Read More
Answer

Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.

I apologize for the inconvenience and am pleased to offer you in compensation a credit note or a free of charge replacement with one of the following anti-aquaporin 4 antibodies :

ab125049 https://www.abcam.com/ab125049,tested inMouse and ICC/IF, but not in IHC-Fr
ab15118 https://www.abcam.com/ab15118, predicted to react with Mouse, tested in WB and ELISA only.
ab81355 https://www.abcam.com/ab81355, predicted to react with Mouse, tested in WBonly.



Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More
Application
Western blot
Sample
Pig Tissue lysate - whole (Jejunum)
Loading amount
60 µg
Specification
Jejunum
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 25 2011

Application
Western blot
Sample
Rat Cell lysate - whole cell (retina)
Loading amount
30 µg
Specification
retina
Gel Running Conditions
Reduced Denaturing (4-20%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Nov 21 2008

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (kidney)
Specification
kidney
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Permeabilization
No
Blocking step
5 minutes of peroxidase block then 10 minutes of protein block. (These are ready-to-use reagents purchased from Dako) as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C

Mr. Antibody Solutions

Verified customer

Submitted Aug 22 2008

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (kidney)
Specification
kidney
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH6.0
Permeabilization
No
Blocking step
5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C

Mr. Antibody Solutions

Verified customer

Submitted Aug 19 2008

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (kidney)
Specification
kidney
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Permeabilization
No
Blocking step
5 minutes of peroxidase block then 10 minutes of protein block. (These are ready-to-use reagents purchased from Dako) as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C

Mr. Antibody Solutions

Verified customer

Submitted Aug 19 2008

21-30 of 30 Abreviews or Q&A

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