Anti-Influenza A Virus Nucleoprotein antibody [AA5H] (ab20343)

phone call: powder instead of liquid in the tube

ANSWER:

Merci pour votre appel l-autre jour.

Nous sommes désolés que vous avez reçu un tube qui ne contenait pas l'anticorps liquide. Est-ce que après centrifugation vous avez récupérez un peu de liquide?

Nous avons fait une investigation de notre part, mais nous n'avons pas trouvé une explication. Nous avons contrôlé tout notre stock aussi - et les tubes en stock sont tous liquides comme attendu.

Je suis désolée qu'il semble que le tube que vous avez reçu était endommagé. Nous allons vous envoyer volontiers un tube de remplacement, si après centrifugation le problème ne s'était pas résolu.

Je me réjouis de votre réponse.

I am trying to do immunoprecipitation with one of your influenza virus antibodies. Anti-Influenza A Virus Nucleoprotein antibody [AA5H] (ab20343) I have read your online protocol for immunoprecipitation http://www.abcam.com/ps/pdf/protocols/Immunoprecipitation%20protocol%20(IP).pdf and I have a doubt, at the point immunoprecipitation: Point 7. Finally, remove the last supernatant and add 25-50 μl of 2x loading buffer. Boil at 95-100°C for 5 minutes to denature the protein and separate it from the protein-A/G beads, then centrifuge and keep the supernatant where the protein is now. Once I do the elution in 2x loading buffer, do I need to further dilute the sample with distilled water 1 in 2 before loading the sample into the gel? Otherwise I will be loading the samples in 2x loading buffer..... Thank you very much for your help in advance!

ANSWER:

You will not need to dilute your sample any further after eluting from the beads. As the protocol says, you can after boiling load your sample directly onto your gel, without it affecting anything. If there is anything else I can help you with, please let me know.

I’ve been using the influenza A nucleoprotein antibody [AA5H] (ab20343) to stain for influenza A PR/8 (H1N1) infected cells. It’s worked great so far. However, I haven’t had success in staining cells (MDCK) with influenza A X-31 (H3N2). According to the specs, AA5H should work with H3N2 subtype. Was wondering if abCam has any information or insight on using AA5H for the X31 strain. I’ve also tried primary antibody 431-FITC on X31; however, I have not had success with that either.

ANSWER:

Thanks for your inquiry.

The monoclonal antibody to Influenza A Nucleoprotein, catalog ab20343, has been tested and is reactive with the following Influenza A strains:

H1N1

A/Puerto Rico/8/34A/USSR/90/77

A/England/333/80

A/Chile/1/83

A/Dunedin/6/83

A/Victoria/7/83

H2N2 A/Ann Arbor/6/60

H3N2

A/Aichi/2/68

A/Port Chambers/1/73

A/Bangkok/2/79

A/Korea/1/82

A/Philippines/2/82

A/Caen/1/84

A/Pennsylvania/27/84

For ab81126 : we used a vaccine as the immunogen of ab81126, and used the recombinant protein NP to screen positive clones. So the antibody of ab81126 can be used to detect the NP protein. We are not sure whether this antibody can react with all strains of influenza A virus, because we haven't done the relating tests.

I hope this was helpful.

Please feel free to contact me if you require further assistance.

Can this antibody detect intact influenza A virus?

ANSWER:

Thank you for your inquiry.

The laboratory confirmed that they have not tested ab20343 against the intact virus.  The following link is to a published reference that describes the development and original characterizaton of this clone.  The paper includes IFA testing of cells infected with viruses reisolated from clinical Influenza A specimens.

http://jcm.asm.org/cgi/reprint/23/2/240

Catalog ab20343, Clone AA5H, is described as Clone A1 in the paper. 

I hope this information helps. Please contact me with any other questions.

My post doc would like to know if you have available the specific recognition sequence for this antibody.

ANSWER:

Thank you for your patience in this matter.

I have received a reply from the originator of ab20343 (Influenza A Virus Nucleoprotein antibody [AA5H]). All the characterization information we have on this product is from the following published reference:

http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=268620&blobtype=pdf

Note that Clone AA5H is Clone A1 in the paper.

I hope this information helps. Please don't hesitate to contact us again if you have further questions.