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  1. Link

    kcnq4-antibody-n436-ab84820.pdf

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Neuroscience Neurotransmission Receptors / Channels Potassium Channels
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Anti-KCNQ4 antibody [N43/6] (ab84820)

  • Datasheet
Reviews (1)Q&A (8)References (2)

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Immunohistochemistry (Frozen sections) - Anti-KCNQ4 antibody [N43/6] (ab84820)
  • Western blot - Anti-KCNQ4 antibody [N43/6] (ab84820)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KCNQ4 antibody [N43/6] (ab84820)

Key features and details

  • Mouse monoclonal [N43/6] to KCNQ4
  • Suitable for: IHC-Fr, IHC-P, WB
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG1

You may also be interested in

Secondary
Product image
Goat Anti-Mouse IgG H&L (HRP) (ab205719)

View more associated products

Overview

  • Product name

    Anti-KCNQ4 antibody [N43/6]
  • Description

    Mouse monoclonal [N43/6] to KCNQ4
  • Host species

    Mouse
  • Tested Applications & Species

    Application Species
    IHC-Fr
    Mouse
    IHC-P
    Human
    WB
    Rat
    See all applications and species data
  • Immunogen

    Fusion protein: AEAPPRRLGL GPPPGDAPRA ELVALTAVQS EQGEAGGGGS PRRLGLLGSP LPPGAPLPGP GSGSGSACGQ RSSAAH, corresponding to amino acids 2-77 of Human KCNQ4 (accession number P56696).

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • Lysate from transfected COS-1 cells transiently expressing KCNQ4.
  • General notes

    The clone number has been updated from S43-6 to N43/6, both clone numbers name the same antibody clone.

     

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    Preservative: 0.09% Sodium azide
    Constituents: 50% Glycerol, PBS
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    N43/6
  • Isotype

    IgG1
  • Research areas

    • Neuroscience
    • Neurotransmission
    • Receptors / Channels
    • Potassium Channels
    • Neuroscience
    • Sensory System
    • Auditory system

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab84820 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
IHC-Fr
Mouse
IHC-P
Human
WB
Rat
All applications
Chimpanzee
Application Abreviews Notes
IHC-Fr
1/1000.
IHC-P
Use at an assay dependent concentration.
WB
1/1000. Predicted molecular weight: 77 kDa.
Notes
IHC-Fr
1/1000.
IHC-P
Use at an assay dependent concentration.
WB
1/1000. Predicted molecular weight: 77 kDa.

Target

  • Function

    Probably important in the regulation of neuronal excitability. May underlie a potassium current involved in regulating the excitability of sensory cells of the cochlea. KCNQ4 channels are blocked by linopirdin, XE991 and bepridil, whereas clofilium is without significant effect. Muscarinic agonist oxotremorine-M strongly suppress KCNQ4 current in CHO cells in which cloned KCNQ4 channels were coexpressed with M1 muscarinnic receptors.
  • Tissue specificity

    Expressed in the outer, but not the inner, sensory hair cells of the cochlea. Slightly expressed in heart, brain and skeletal muscle.
  • Involvement in disease

    Defects in KCNQ4 are the cause of deafness autosomal dominant type 2A (DFNA2A) [MIM:600101]. DFNA2A is a form of sensorineural hearing loss. Sensorineural deafness results from damage to the neural receptors of the inner ear, the nerve pathways to the brain, or the area of the brain that receives sound information.
  • Sequence similarities

    Belongs to the potassium channel family. KQT (TC 1.A.1.15) subfamily. Kv7.4/KCNQ4 sub-subfamily.
  • Domain

    The segment S4 is probably the voltage-sensor and is characterized by a series of positively charged amino acids at every third position.
    The A-domain tail carries the major determinants of channel assembly specificity. Its coiled-coil region is Four-stranded.
  • Cellular localization

    Basal cell membrane. Situated at the basal membrane of cochlear outer hair cells.
  • Target information above from: UniProt accession P56696 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 456800 Chimpanzee
    • Entrez Gene: 9132 Human
    • Entrez Gene: 60613 Mouse
    • Entrez Gene: 298496 Rat
    • Omim: 603537 Human
    • SwissProt: P56696 Human
    • SwissProt: Q9JK97 Mouse
    • SwissProt: Q9JK96 Rat
    • Unigene: 473058 Human
    • Unigene: 249977 Mouse
    • Unigene: 144875 Rat
    • Unigene: 225305 Rat
    see all
  • Alternative names

    • DFNA 2 antibody
    • DFNA2 antibody
    • KCNQ 4 antibody
    • Kcnq4 antibody
    • KCNQ4_HUMAN antibody
    • KQT like 4 antibody
    • KQT-like 4 antibody
    • KV7.4 antibody
    • Potassium channel KQT like 4 antibody
    • Potassium channel subunit alpha KvLQT4 antibody
    • Potassium voltage gated channel KQT like protein 4 antibody
    • Potassium voltage gated channel KQT like subfamily member 4 antibody
    • Potassium voltage gated channel subfamily KQT member 4 antibody
    • Potassium voltage-gated channel subfamily KQT member 4 antibody
    • Voltage gated potassium channel subunit Kv7.4 antibody
    • Voltage-gated potassium channel subunit Kv7.4 antibody
    see all

Images

  • Immunohistochemistry (Frozen sections) - Anti-KCNQ4 antibody [N43/6] (ab84820)
    Immunohistochemistry (Frozen sections) - Anti-KCNQ4 antibody [N43/6] (ab84820)

    ab84820 staining KCNQ4 in Mouse brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections).

  • Western blot - Anti-KCNQ4 antibody [N43/6] (ab84820)
    Western blot - Anti-KCNQ4 antibody [N43/6] (ab84820)
    All lanes : Anti-KCNQ4 antibody [N43/6] (ab84820)

    Lane 1 : Molecular weight marker
    Lane 2 : Mixed cell lysates prepared from rat tissues Brain, Heart, Kidney, Liver, Lung, Pancreas, Skeletal Muscle, Spleen, Testes, Thymus, and H9C2 cells at 10 µg

    Predicted band size: 77 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KCNQ4 antibody [N43/6] (ab84820)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KCNQ4 antibody [N43/6] (ab84820)
    ab84820 staining KCNQ4 in human hippocampal tissue by IHC-P (Bouin's fixed paraffin embedded).

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (2)

    Publishing research using ab84820? Please let us know so that we can cite the reference in this datasheet.

    ab84820 has been referenced in 2 publications.

    • Brueggemann LI  et al. Differential protein kinase C-dependent modulation of Kv7.4 and Kv7.5 subunits of vascular Kv7 channels. J Biol Chem 289:2099-111 (2014). WB, ICC/IF . PubMed: 24297175
    • Udagawa T  et al. Inwardly rectifying potassium channel Kir4.1 is localized at the calyx endings of vestibular afferents. Neuroscience 215:209-16 (2012). PubMed: 22546335

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-9 of 9 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-KCNQ4 antibody [S43-6]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: RT°C
    Sample
    Mouse Cell (NIH3T3 TRANSFECTED KCNQ4 CELL)
    Specification
    NIH3T3 TRANSFECTED KCNQ4 CELL
    Permeabilization
    Yes - triton X100 0.5%
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 31 2014

    Question

    Thank you for your troubleshooting. I think the best course of action would be try a different antibody as I am working to a strict time limit and I do not have alot of time to optimise each condition.



    I have looked through the antibodies and I have found a different one which may be more suitable. The product code is

    ab56456.



    I will be more than happy to give feedback in exchange for the new antibody.



    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on May 28 2012

    Answer

    Thank you for your answer and for your cooperation.

    I am sorry that the ab84820 required more optimisation than expected and for the inconvenience this has caused.As requested, I agreed to send a free alternative antibody in exchange for the feedback via an Abreview for both antibodies. I have issued now this free of charge replacement with the alternative antibody ab56456 with the order number 1096821.

    To check the status of the order please contact our Customer Service team and reference this number.

    We look forward hearing back from you andwish you the best of luck with your research.

    Read More

    Abcam Scientific Support

    Answered on May 28 2012

    Question

    Thank you for your email. I have addressed the issues you have mentioned with a post-doc in the lab and she had optimised our western blotting protocol as per the manufactures recommendations of membrane blocking. She has also performed membrane blocking with BSA and milk and there is no difference between BSA and milk or between BSA and milk and methanol.

    Would it be possible for you to send an antibody raised against a different epitope?

    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on May 25 2012

    Answer

    Thank you for your reply.

    We can send you a new antibody in exchange for your feedback, however I am not sure whether this will resolve your question.

    I have discussed this case also with a colleague of mine. Indeed, we both think that the results obtain seem to correlate and that with a new antibody we would expect the same results. The reason why we think the results to correspond to what is expected are:
    1.) The results improved upon reduction of the heating, which correspond to the expected behaviour of this target protein.
    2.) The results show some duplicate band, which would correspond to the two isoforms as described on SwissProt (http://www.uniprot.org/uniprot/P56696),
    3.) I agree that the observed molecular weight is not as expected, however,as mentioned in my previous emails, Ithink it would be very usefulto compare the compatibility of the buffer and molecular weight marker. Different molecular weight marker migrate different in different buffer systems. Variations of 15% in the expected molecular weight can be seen. See the images of our MW marker as an example: https://www.abcam.com/Prism-Ultra-Protein-Ladder-10-245-kDa-ab116028.html

    Please let me know what you think about these points. So far, we had no other negative feedback on this antibody neither. I can provide you with an alternative antibody if you provide us feedback on both antibodies, used in the same conditions, via an Abreview (https://www.abcam.com/abreviews). We do have several other antibodies against KCNQ4 which have been tested in Western blot and on human cells. Please see here the links to their datasheets:
    https://www.abcam.com/KCNQ4-antibody-ab56456.html (or use the following: https://www.abcam.com/KCNQ4-antibody-ab56456.html).
    https://www.abcam.com/KCNQ4-antibody-ab65797.html (or use the following: https://www.abcam.com/KCNQ4-antibody-ab65797.html).
    https://www.abcam.com/KCNQ4-antibody-ab82096.html (or use the following: https://www.abcam.com/KCNQ4-antibody-ab82096.html).
    https://www.abcam.com/KCNQ4-antibody-ab110373.html (or use the following: https://www.abcam.com/KCNQ4-antibody-ab110373.html).

    Thank you very much for your cooperation. Please let me know if you have any questions. I look forward to hear your opinion on the above mentioned points and how you would like to proceed.

    Read More

    Abcam Scientific Support

    Answered on May 25 2012

    Question

    Thank you. Regarding membrane blocking, we do not incubate the membrane before primary antibody incubation. As mentioned before, we submerge the membrane in methanol for 15 seconds.

    I am out of the office for a week so I will have answers to your other suggestions when I get back.

    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on May 22 2012

    Answer

    I just write you to inquire whether you have yet had the opporunity to look into my comments/suggestions? Please let me know also if you have any questions or concerns - I would be happy to discuss them with you. If the problem has been solved, I would also appreciate if you can let me know, so I can update our records accordingly.

    Here is also a link to our online Western blot protocol, where each step of theWestern blotprocedureis described in detail: https://www.abcam.com/index.html?pageconfig=resource&rid=11375
    Blocking the membranes with proteins, such as milk or BSA, is very important to reduce the background. Also, changing from one of those reagents to the other one can significantly improve the results (e.g. Western blot image on our datasheet for ab9385 https://www.abcam.com/HRP-GAPDH-antibody-Loading-Control-ab9385.html (or use the following: https://www.abcam.com/HRP-GAPDH-antibody-Loading-Control-ab9385.html).).

    I am looking forward to hear back from you. Please do not hesitate to contat us also should you have any other question.

    Read More

    Abcam Scientific Support

    Answered on May 22 2012

    Question

    Thank you for your response.

    Regarding the blot images I included in the questionnaire, the blot on the left was run with cell lysates boiled at 95 degrees. The blot on the right was run with tissue lysate boiled at various temperatures from no boiling to 95degrees.

    All tissue/cell samples are human in origin so I do not think it is appropriate to speculate about mouse proteins at this stage.

    The initial blocking is done using methanol for 15 seconds. This seems to work for all members in the lab for a wide range of antibodies and tissue samples.

    I hope this information is helpful to your investigation and can further define the problem.

    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on May 03 2012

    Answer

    Thank you for your reply.

    I am sorry for the confusion of the origin of the samples. Indeed, as you are using human samples, there are two isoformes described. One is at 71kD and the other one is at 77kD. Please see also this link to SwissProt for the details: http://www.uniprot.org/uniprot/P56696
    As I can not read the molecular weight marker annotation on your Western blot, please check whether this could corrspond to the two bands you are observing on the WB on the right side.

    Please check also whether the protein ladder is compatible with the migration buffer. Indeed, depending on the buffer, the protein ladders can migrate at different molecular weights. Please see the images on one of our protein ladders for illustration: https://www.abcam.com/Prestained-Protein-Ladder-Broad-molecular-weight-10-245-kDa-ab116028.html (or use the following: https://www.abcam.com/Prestained-Protein-Ladder-Broad-molecular-weight-10-245-kDa-ab116028.html).

    When I was asking about the blocking of the membrane, I ment the protein blocking - before performing the immunostaining. Do you not incubate the membranes in a protein solution with either milk or BSA before incubating with the primary antibody dilution? As you are using BSA in the antibody dilution, I would think that you are using also a BSA blocking step prior to the antibody incubation?

    Please let me know your feedback. If these points do not solve the problem, I will be pleased to continue to investigate the problem. If the antibody is not working, please be assured that is covered by our Abpromise guarantee to work as stated on the datasheet and we would therefore replace it (if purchased within the last 6 months).

    Unfortunately, the laboratory has no other images as the one on the datasheet, nor do we have tubes of the immunogen fragement available. I am sorry.

    I am looking forward to hear back from you.

    Read More

    Abcam Scientific Support

    Answered on May 03 2012

    Question


    Please find attached a copy of my completed questionnaire.



    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on May 02 2012

    Answer

    Thank you for taking time to complete our questionnaire. I am sorry that 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.

    Thank you very much for the two images as well. Can you explain to me what the difference is between the blots on the left and the blot on the right?

    As discussed, I have contacted the laboratory for more information and explaining the problem.The laboratory replied thatit sounds like the band is in the right area, but that a doublet would generally mean a small difference in size.It could be a deletion, or proteolysis fragment.The laboratory has not seen this, but that is not to say it cannot occur, as this isvery much an evolving area.The work of the laboratorywas on rat brain. In summary, the laboratory head thinks thatthe fact to havea doublet could either be due to a preparation artifact, proteolysis fragment or possibly a generic variation, deletion, pseudogene etc.

    Indeed, there is no described isoform to my knowledge in mouse, however in human there are two isoformes, one at 77kD and one at 71kD. It might well be therefore, that the mouse isoforme still needs to be discovered? http://www.uniprot.org/uniprot/P56696

    In order also to exclude any artifact, have you checked whether any background banding could be due to the secondary antibody? This could easily be checked by running the same samples and the same blot just without the primary antibody.

    Can you please also let me know how the membranes have been blocked, have you used milk or BSA? Often changing the blocking buffer can also improve the signal.

    Please let me know your thoughts. I am still following up on the possiblity of the blocking peptide and the images and will get back to you in this regards.

    As mentioned before, in the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

    I hope this information is helpful, and I thank you for your cooperation.

    Read More

    Abcam Scientific Support

    Answered on May 02 2012

    Question

    Phone call: uses human uterus and obtaines mutliple bands - also a doublet around 80kD. has been in contact with us previsouly

    Read More

    Abcam community

    Verified customer

    Asked on May 01 2012

    Answer

    Thank you for calling us.

    As we discussed on the phone, I have contacted thelaboratory for the additional information. I am also attaching the discussed questionnaire so that we can gather further information regarding the samples tested and the protocol used. This will enable us to look with more details into the results you obtained and try to troubleshoot them.

    If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

    I look forward to receiving your reply.

    Read More

    Abcam Scientific Support

    Answered on May 01 2012

    Question

    Please provide WB protocol used with ab84820.

    Multiple bands are observed, what troubleshooting tips you suggests.

    Read More

    Abcam community

    Verified customer

    Asked on Mar 12 2012

    Answer

    Thank you very much for your email.

    I have now received an email from lab. What my colleague has suggested is attached below;

    Multiple bands are very common with ion channels in general. Proteolysis is very common – it has to do with liberating the channels from the membranes – the protective barrier against the proteases is lost. What I would suggest is two-fold. The use of proteolysis inhibitor cocktails when preparing the lysate, and secondly, do not boil it or heat to 90°C when prepping it to run on SDS. Keep all conditions the same but then instead heat at 37°C for 15 minutes instead. That worked for us, although the outcome is still of course dependent on the initial proteolysis if there is any. Other than that any western protocol should work just fine. Proteases tend to be very heat resistant (they can work very well at 55°C+), and because (all) the reactions are still temperature dependent for the short time they are active at the high temperature they can inflict a high level of degradation.

    I hope this information will be helpful. Should you have any other inquiry please do not hesitate to contact me.

    Read More

    Abcam Scientific Support

    Answered on Mar 12 2012

    Question

    Please provide WB protocol used with ab84820.

    Read More

    Abcam community

    Verified customer

    Asked on Mar 09 2012

    Answer

    I can confirm that I have send an email to my colleaguein lab and requested the productspecific WB protocol; I will be able to forward the same once I get a reply from them.

    Many thanks for having patience.

    Read More

    Abcam Scientific Support

    Answered on Mar 09 2012

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