Anti-Influenza A Virus M2 Protein antibody [14C2] (ab5416)

Reviews (2)Q&A (17)References (46)

Overview

  • Product name

    Anti-Influenza A Virus M2 Protein antibody [14C2]
    See all Influenza A Virus M2 Protein primary antibodies

  • Description

    Mouse monoclonal [14C2] to Influenza A Virus M2 Protein

  • Host species

    Mouse

  • Tested applications

    Suitable for: Inhibition Assay, Flow Cyt, ICC/IF, ICC, IP, WBmore details

  • Species reactivity

    Reacts with: Influenza A

  • Immunogen

    Tissue, cells or virus corresponding to Influenza A Virus M2 Protein. Full length M2 protein from A/WSN/33-infected CV1 cell lysate.

  • Epitope

    Detects the N-terminal of the Influenza A Virus M2 Protein.

  • Positive control

    • ICC/IF: Infected MDCK cells.

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: 0.05% Sodium azide
    Constituents: 99% PBS, 0.1% BSA
  • Concentration information loading...

  • Purity

    Protein G purified

  • Primary antibody notes

    Influenza A virus is an enveloped virus encoding 10 polypeptides. RNA segment 7 encodes for two proteins: M1 (matrix 1) and M2 (matrix 2). M1 protein is encoded by an mRNA that is colinear, while M2 protein is synthesized from spliced mRNA. M2 protein is a transmembrane protein composed of three Domains: 1) 24 residues representing the N-terminal region, 2) 19 hydro-phobic residues that serve as a membrane anchor, and 3) 54 residues near the C-terminal in the cytoplasmic domain. The M2 protein has been found to play a role in Influenza replication and assembly of virion particles. Further experimentation has demonstrated that this protein is an acid-activated ion channel for virus replication.

  • Clonality

    Monoclonal

  • Clone number

    14C2

  • Isotype

    IgG1

  • Light chain type

    kappa

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab5416 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
Inhibition Assay Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF Use a concentration of 1 µg/ml.
ICC Use a concentration of 1 µg/ml.
IP Use a concentration of 10 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 15 kDa.

Target

  • Relevance

    The Matrix protein M2 forms a protons channel. When the environmental pH is lower than a threshold, the M2 channel is activated and selectively transports protons accross the membrane from the extracellular side to the cytoplasmic side. It is crucial for the uncoating process. When the virion is internalized into the endosome the channel can acidify the virion interior, promoting the dissociation of the viral matrix protein (M1) from the ribonucleoprotein (RNP) thus allowing the transport of the RNP from the virion into the cell’s nucleus. For some influenza virus subtypes, the M2 channel can elevate the intravesicular pH of the trans Golgi network, preventing the viral protein haemagglutinin, which is transported to the cell surface through the trans Golgi network, from incorrect maturation in an otherwise low pH environment.

  • Cellular localization

    Virion membrane. Apical cell membrane; Single-pass type III membrane protein.

  • Database links

    • Alternative names

      • Influenza A virus matrix protein M2 antibody
      • M antibody
      • M2 antibody
      • Matrix protein 2 antibody
      • Membrane ion channel M2 antibody
      • Membrane protein M2 antibody
      • Proton channel protein M2 antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-Influenza A Virus M2 Protein antibody [14C2] (ab5416)
      Immunocytochemistry/ Immunofluorescence - Anti-Influenza A Virus M2 Protein antibody [14C2] (ab5416)
      Immunofluorescence staining of infected MDCK cells using ab5416.

    References

    This product has been referenced in:

    • Ma X  et al. Transcription Factor EB Activation Rescues Advanced aB-Crystallin Mutation-Induced Cardiomyopathy by Normalizing Desmin Localization. J Am Heart Assoc 8:e010866 (2019). Read more (PubMed: 30773991) »
    • Yao Y  et al. Protection against homo and hetero-subtypic in?uenza A virus by optimized M2e DNA vaccine. Emerg Microbes Infect 8:45-54 (2019). Read more (PubMed: 30866759) »
    See all 46 Publications for this product

    Publishing research using ab5416? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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

    1-10 of 19 Abreviews or Q&A

    Question

    Please send me a quote for 3 vials of BSA and azide free influenza A antibody.

    Best regards

    Read More
    Answer

    Thank you very much for your patience.

    Finally, I have a price quote and custom made confirmation from laboratory. We are happy to provide this product at lower cost than ab5416. The price would be £194 per vial or £582 for 3 vials of this product, excluding shipping charges which will be £16 for UK and 21 Euros for Germany.

    Could you let me know if you want to go ahead with the order? I will then be happy to publish the datasheet of this custom made product.

    Please note for proper quotation I would need the billing and shipping address.

    I will look forward to hearing from you soon.

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    Question

    Please send me a quote for 3 vials of BSA and azide free influenza A antibody.

    Best regards

    Read More
    Answer

    Thank you for your email.

    I have sent a request to laboratory for quote; I will send you the details soon.

    Many thanks for having patience!

    Read More

    Question

    I would like to buy two vials of the BSA and azide free monoclonal antibody against the Influenza A M2 protein (see below).

    vial volume: 100µL

    antibody concentration: 1mg/mL stored in PBS

    Is it possible to get an offer?

    Thank you!

    Best regards

    Read More
    Answer

    Thank you for your email.

    I just spoken to my colleague in laboratory; they have suggested that in order to justify the production cost for making it BSA and azide free at least 3 vials should be ordered. Could you let me know if you will buy 3 vials and then I will send you a quote.

    I will look forward to hearing from you soon. Let me know if you have any questions and concerns.

    Read More

    Question

    I bought the antibody (ab5416) month ago. But the antibody is stored in a 1mg/mL BSA solution. Do you have this antibody also in a BSA free solution?

    In an earlier search I found the ab96689.

    Best regards!

    Read More
    Answer

    Thank you for contacting us.

    We can provide this product, BSA and azide free as a special request. Could you let me know how many vials you would like to buy?

    Looking forward to hearing from you soon.

    Read More

    Question

    Customer kindly called to inquire whether this product would recognize the Influenza A PR8 strain.

    Read More
    Answer

    Thank you for contacting us.

    Our Influenza A M2 Monoclonal Antibody (14C2) - product #ab5416 was shown to detect influenza A virus M2 strain A/PR/8/34 in the following reference:



    Zebedee SL, Lamb RA"Influenza A virus M2 protein: monoclonal antibody restriction of virus growth and detection of M2 in virions."J Virol. 1988 Aug;62(8):2762-72.
    (http://www.ncbi.nlm.nih.gov/pubmed/2455818)

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

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

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    Question

    Thank you very much for your help!

    1. I measured the OD280 of the various steps from my sample using the Nandrop ND1000. I did not dilute the samples (except for the original antibody stock) prior to measuring.
    Raw A280 values:
    Original antibody solution (50% concentration) = 0.712
    Unbound fraction = 1.46
    Wash1 = -0.022
    Wash2 = -0.014
    Wash3 = 0.022
    Wash4 =0.015
    Elution1 =0.069
    Elution2 = -0.013
    Elution3 = 0.038
    Elution4 = -0.032

    2. Please confirm the volumes used. When calculating recoveries and absolute amounts of material we need to know the volume as well as the concentration.
    For the volumes used, I used 100uL of the antibody and added 10uL of binding buffer to it. This was mixed and applied to the resin in the spin column. The resin itself was reconstituted using 0.5mL of wash buffer. The mixture was incubated at 25C in a thermomixer for 1 hour.
    For the wash steps, I used 0.5mL wash buffer 4 times, and spun down 30s as stated in the protocol
    For the elution steps, I added 100uL of elution buffer to the spin catridge containing the resin and protein, and incubated this at 25C for 2 min, in a thermomixer. This was then spun down for 30 sec. I then added 25uL of the neutralization buffer to the eluate, and stored this on ice for the rest of the procedure. I repeated this elution step 4 times.

    Thank you for your help.
    Regards,

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    Answer

    Thank you foryour reply and forkindly providing the requested information.

    I have investigated this with our source for the antibody and reviewed the details. It is worth consideringthat the Nandrop will be unable to distinguish between IgG and BSA. Also the OD280 nm values are very low and a little variable.It is difficult to draw an absolute conclusion but this does indicate that the antibody did not bind to the column. However, this can be confirmed with further investigation.

    Unfortunately, experience has shown that the measurement of antibody recovery using the nanodrop is not always very accurate.We would therefore recommendto confirmrecovery of the antibody by an alternative method. For example, a 1D non reducing gel would provide an indication of the amount of antibody and its quality. There shouldbe asingle antibody band of approximately 160 kDa. The intensity of this band will be related to the amount of antibody recovered.

    If the recovery is in fact found to be low then it means that for some reason the antibody has failed to bind to the resin. The antibody will therefore be in the flow through. Again a 1D non reducing gel of the flow through sample could confirm this (the material will have two bands IgG and BSA). In this case you could repeat the binding to the resin after adding a second volume of binding buffer. The second addition of binding buffer will make sure that the pH of the antibody is definitely correct (pH 8.0) and that the ionic strength of the antibody buffer is sufficiently raised to promote binding. The kits are provided with sufficient excess buffers to enablethis procedure. The addition of more binding buffer has proved very successful in the past when dealing with troublesome mouse antibodies.

    I hope this will be helpful to you. Please dolet me know how you get on. If you still have some concerns, please do not hesitate to contact me with further information and details of how you would like to proceed.

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    Question

    Hi, I would like to know whether this antibody is also a neutralizing antibody,i.e. preventing cells from Flu virus infection once ab5416 is added to the cell culture. Thx

    Read More
    Answer

    Thank you for contacting Abcam regarding this,
    I have had the chance to speak with the lab about your question. We have not tested whether this antibody can be used as a neutralizing antibody. The authors in the following paper used this antibody in a blocking assay. They found out that "The level of M2 surface expression in A/Udorn virus-infected MDCK cells was found to be reduced to ∼60% of control levels in cells incubated with the 14C2 antibody. In contrast, M2 surface expression levels in A/WSN virus-infected cells were decreased by only ∼5-15%, and A/WSN virus assembly appeared to be unaffected by the M2 antibody treatment." It appears that the blocking is also virus strain dependent.

    http://www.sciencedirect.com/science/article/pii/S0042682285714985

    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.

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    Question

    I am using a 1% concentration of Sodium Deoxycholate in a buffer composed of 10 mM Tris, 150 m M NaCl,PH 8. SDO forms a viscous gel and precipitates!!!

    To my knowledge, SDO precipitates in a PH lower than 7 but PH of my buffer is 8.

    I would be much grateful if you help me with this problem. I have heard about NaCl concentrations above 0.1% which makes SDO insoluble. Is that correct?

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    Answer

    I would suggest to prepare buffers that are recommended in the following Western Blotting protocol : https://www.abcam.com/index.html?pageconfig=resource&rid=13045 .

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    Question

    I have a question regarding M2 Antibody I have ordered to you... Anti-Influenza A Virus M2 Protein antibody [14C2] (ab5416) Actually I want to use this antibody against my virus which is Influenza A Virus (A/New Jersey/8/1976(H1N1) But this antibody is against Influenza A virus (A/WSN/1933(H1N1) Could you please let me know if it is matched with my target protein from conformation point of view? because I have used it for FITC and HRP conjugated antibodies, but no result detected... I think this primary is not match with my target!!! and also ask this question from Abcam Company: If they can suggest or provide me any suitable antibody for this detection? Thank you very much .

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    Answer

    Thank you very much for your inquiry. I have contacted the laboratory for information on the ab5416. However the laboratory has not tested this antibody neither on your strain. I did a sequence alignement of the immunogen and your sequence. Please see the results below. Indeed, the alignement is very good. However as this is a monoclonal antibody, we do not know the epitope and can therefore not guarantee crossreactivity. http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111223-121219-0395-63524695-pg We do have two other antibodies against the Influenza A Virus M2 protein, the ab56086 and the ab94925. I have already done the alignement of the immunogen of ab56086 with your protein of interest. The overlapp is 100%. I would therefore expect that this antibody does recognize your protein as well. xxx I am checking with the laboratory for the alingment of ab94925 with your protein (http://www.uniprot.org/uniprot/A5GX42) and will get back to you. If the customer would like to test the ab56086 on the Jersey Strain, I would be pleased to provide also a testing discount (www.abcam.com/collaborationdiscount). Please let me know if you would like to know more about this testing discount. Please do not hesitate to contact me, should you or the customer have any other question or concern.

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    Question

    Can you provide the exact epitope sequence in the N terminal region where this product, Influenza A Virus M2 Protein antibody [14C2] (MA1-082), detects? Also, does this product detect all sub-types (e.g. H1N1, H5N1)?

    Read More
    Answer

    Thank you for your enquiry. This product was created against whole M2 protein purified from A/WSN/33-infected CV1 cell lysate. Unfortunately, the exact epitope for this polyclonal antibody is not known at this time. The antibody was not tested against all subtypes, but the M2 protein should be identical in the influenza A virus subtypes.

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    1-10 of 19 Abreviews or Q&A

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