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
  • 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.
  • 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

  • Immunofluorescence staining of infected MDCK cells using ab5416.

References

This product has been referenced in:
  • Tutykhina I  et al. Vaccination potential of B and T epitope-enriched NP and M2 against Influenza A viruses from different clades and hosts. PLoS One 13:e0191574 (2018). Read more (PubMed: 29377916) »
  • Opriessnig T  et al. Comparison of the efficacy of a commercial inactivated influenza A/H1N1/pdm09 virus (pH1N1) vaccine and two experimental M2e-based vaccines against pH1N1 challenge in the growing pig model. PLoS One 13:e0191739 (2018). WB . Read more (PubMed: 29381710) »
See all 33 Publications for this product

Customer reviews and Q&As

1-10 of 19 Abreviews or Q&A

Question
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.

Read More

Question
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

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

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

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

Read More

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.

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.

Read More

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 .

Read More

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.

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.

Read More

1-10 of 19 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up