Anti-Dengue Virus NS1 glycoprotein antibody [DN3] (ab41616)

Thank you for contacting us.

DN3 is just a clone name. The antibody is reactive to all serotypes.

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

Thank you for your message which has been forwarded to me as my colleague Anja is currently out of the office.

We appreciate the feedback from the customer, thank you for sending this information.

In order to obtain the discount, the customer will need to submit this informationan Abreview. They can do this by clicking on the Abreviews tab on the datasheet, then click on the link to 'submit an Abreview'.

Please click the 'Submit and Abreview' link on the following page:

https://www.abcam.com/Dengue-Virus-NS1-glycoprotein-antibody-DN3-ab41616-reviews.html

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this will be helpful. If you have any further questions regarding submission of the Abreview, please do not hesitate to letme know.

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

I would like to reassure you that this antibody is tested and covered by our guarantee for WB. Before deciding how to proceed, I would like to investigate this particular case further for you, and also obtain some further information for our quality records to determine what may be the cause of the problem.

In order to do this, I have enclosed a questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. I have completed the sections for which information has already been provided.

It would also be beneficial if you are able toconfirm:

1. What other samples have been tested with this antibody? Has it detected endogenous protein in any samples? I can suggest it would be beneficial to try this if possible.

2. Has the protein been stained with comassie on an SDS gel? Is it intact and running at the correct molecular weight?

In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.


Order Details
Antibody code:

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


Amount of protein loaded


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

Thank youand your customer very much for your interest in anti dengue virusNS1 antibodies.

I can confirm that we have three in our catalog: ab41490, ab41616 and ab41623.

To our knowledge, none of these antibodies has been tested in ELISA yet.

Therefore, I can offer a discount off a future purchase if your customerbuysone of them now, tests it in ELISA and submits feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your customer'sfeedback. The discount would be to the value ofthe antibodyand can be used for the next order of any product.

If your customer is interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that your customer ( please include theire-mail address)would like to proceed and confirm which one of these antibodiesyour customer would like to testin ELISA . I will then send a discount code. This code must be issued before purchasingthe antibodyso please wait for my reply before ordering.

2. Purchasethe antibodyeither by phone, fax, or online (www.abcam.com).

3. Test it in ELISA.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your customer's new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any product ordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if teh antibody turns out to be unsuitable for ELISA, your customerwill still receive the discount ontheir next purchase after your Abreview has been submitted.

Please let me know if you or your customer have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Thank youfor your inquiry.

I can confirm that we do have three anti NS1 antibodies for Dengue virus in our catalog..

Isotype control are not included and have to be purchased separately.

ab41616 (Anti-Dengue Virus NS1 glycoprotein antibody [DN3]):

https://www.abcam.com/Dengue-Virus-NS1-glycoprotein-antibody-DN3-ab41616.html (or use the following: https://www.abcam.com/Dengue-Virus-NS1-glycoprotein-antibody-DN3-ab41616.html).

Heaton NS et al. Dengue virus nonstructural protein 3 redistributes fatty acid synthase to sites of viral replication and increases cellular fatty acid synthesis. Proc Natl Acad Sci U S A 107:17345-50 (2010). ICC/IF. PubMed: 20855599

Isotype control: ab18447 (Mouse IgG1, kappa monoclonal [MG1-45] - Isotype control)
https://www.abcam.com/Mouse-IgG1-kappa-monoclonal-MG1-45-Isotype-Control-BSA-and-Azide-free-ab18447.html (or use the following: https://www.abcam.com/Mouse-IgG1-kappa-monoclonal-MG1-45-Isotype-Control-BSA-and-Azide-free-ab18447.html).

ab41623 (Anti-Dengue Virus NS1 glycoprotein antibody [DN2]):

https://www.abcam.com/Dengue-Virus-NS1-glycoprotein-antibody-DN2-ab41623.html (or use the following: https://www.abcam.com/Dengue-Virus-NS1-glycoprotein-antibody-DN2-ab41623.html).

Smith KM et al. Structural mutants of dengue virus 2 transmembrane domains exhibit host-range phenotype. Virol J 8:289 (2011). IF. PubMed: 21658241

Isotype control: ab18447 (Mouse IgG1, kappa monoclonal [MG1-45] - Isotype control)
https://www.abcam.com/Mouse-IgG1-kappa-monoclonal-MG1-45-Isotype-Control-BSA-and-Azide-free-ab18447.html (or use the following: https://www.abcam.com/Mouse-IgG1-kappa-monoclonal-MG1-45-Isotype-Control-BSA-and-Azide-free-ab18447.html).

ab41490 (Anti-Dengue Virus NS1 glycoprotein antibody [DN1]):

https://www.abcam.com/Dengue-Virus-NS1-glycoprotein-antibody-DN1-ab41490.html (or use the following: https://www.abcam.com/Dengue-Virus-NS1-glycoprotein-antibody-DN1-ab41490.html).

Currently we do not know of any references using this antibody.

Isotype control: ab18449 (Mouse IgG2a, kappa [MOPC-173] (low endotoxin) - isotype control)
https://www.abcam.com/Mouse-IgG2a-kappa-monoclonal-MOPC-173-Isotype-Control-Low-endotoxin-Azide-free-ab18449.html (or use the following: https://www.abcam.com/Mouse-IgG2a-kappa-monoclonal-MOPC-173-Isotype-Control-Low-endotoxin-Azide-free-ab18449.html).

We do not have set expiration dates for our products. Most antibodies are stable and can last for anywhere from a few months to several years if stored properly, so we strongly recommend that you follow the storage instructions on the datasheet for the antibody you purchased. These conditions will vary among our antibodies, therefore, it is important to verify the storage conditions for each of our products when you receive them. We guarantee all of our products to work for at leastsix monthfrom the date of purchase when stored correctly.

For more information on antibody storage and stability, please visit our Antibody Storage Guide on our Protocols and Troubleshooting Tips page (https://www.abcam.com/index.html?pageconfig=popular_protocols).

We do not sell directly to Pakistan. Please contact our trusted distributor for pricing information:

Friends Scientific Corporation
3-B Ali Continental,
1-Mozang Road
Lahore
54000
Pakistan

Tel 4237236176 Fax 4237321500 Email mailto:friends1@wol.net.pk

I hope this information is helpful and wish you good luck with your experiments.

Gracias por tu respuesta.
He consultado esta información con mis compañeros del laboratorio.
La glicoproteína (ab64456) contiene toda la secuencia excepto el dominio transmembranal. Además contiene 2x6 His tags, uno localizado en el extremo N terminal y el otro en el C terminal.
Respecto al anticuerpo (ab41616) me comentan que el dímero del virus Dengue NS1 está presente en las muestras sin calentar (forma sin reducir), y por efecto del calor se convierte al monómero. El epítopo es por tanto conformacional. En cualquier caso imagino que para llevar a cabo el ensayo ELISA no usáis muestras reducidas. Si tenéis la oportunidad os aconsejaría que probarais la combinación de proteína /anticuerpo mediante WB usando las muestras sin reducir, para comprobar si el anticuerpo realmente está detectando la proteína.
El epítopo del anticuerpo no se ha mapeado, por tanto no puedo saber si reconoce el extremo N terminal o C terminal de la proteína donde se localizan los His tags.
En cualquier caso, como no hay más Dengue Virus NS1 glicoproteínas en el catálogo, puedo mandaros de reemplazo otro primario contra la misma, esperando que esta vez si la reconozca. Estas son las alternativas:
https://www.abcam.com/Dengue-Virus-NS1-glycoprotein-antibody-DN1-ab41490.html
https://www.abcam.com/Dengue-Virus-NS1-glycoprotein-antibody-DN2-ab41623.html]
Espero vuestra respuesta para saber cómo proceder y qué solución os podemos dar.

Thank you for your inquiry.

ab41616 is a mouse monoclonal IgG1 with a kappa light chain, un-conjugated.

Therefore a isotype control is needed that is also un-conjugated and of the IgG1 Isotype (preferably with a kappa light chain). Here are three possible antibodies:

ab18447
https://www.abcam.com/Mouse-IgG1-kappa-monoclonal-MG1-45-Isotype-Control-BSA-and-Azide-free-ab18447.html (or use the following: https://www.abcam.com/Mouse-IgG1-kappa-monoclonal-MG1-45-Isotype-Control-BSA-and-Azide-free-ab18447.html).

ab18443
https://www.abcam.com/Mouse-IgG1-kappa-monoclonal-MOPC-21-isotype-control-ab18443.html (or use the following: https://www.abcam.com/Mouse-IgG1-kappa-monoclonal-MOPC-21-isotype-control-ab18443.html).

ab91353
https://www.abcam.com/Mouse-IgG1-Kappa-Monoclonal-B116-Isotype-Control-ab91353.html (or use the following: https://www.abcam.com/Mouse-IgG1-Kappa-Monoclonal-B116-Isotype-Control-ab91353.html).

I hope this information is helpful and wish your customer good luck with their research.

Thank you for contacting us.
I am sorry to hear you are having problems with one of our products. I appreciate the details provided. I am afraid that product ab41616 is not guaranteed to work in ELISA. We have tested this antibody in WB and ICC, but we are not aware of its suitability for ELISA assays.
As far as I understand you are performing an indirect ELISA, coating your plate with the antigen ab64456 and then capturing it with the monoclonal antibody, using ab20043 (secondary) to detect your primary, is that correct? It could well be the primary antibody is not recognising ab64456 as it may not work for this application.
I can also confirm the peptide ab64456 is tagged with 6 histidines. It is also possible the antibody is not detecting this protein if the epitope it recognizes is masked with the histidine tags.
If you wish to I can try to find out the epitope targeted by ab41616 and check whether it is comprised within the tag region.
Regarding the failure of ab1187 (Anti-6X His tag® antibody (HRP)) to detect the glycoprotein, it could be due to the presence of sodium azide to preserve the protein. The antibody is linked to HRP and this enzyme is very sensitive to sodium azide.
In any case, as all of our products are covered by our Abpromise guarantee, if you wish to receive a replacement for ab64456, I will be more than happy to send it to you.
However, if you want to receive the same protein, you may still experience the same problems. In order to target this protein, I can try to find within our catalogue a primary against it that does not detect the epitope within the tagged sequence, or any other solution you may prefer.
I look forward to receiving your reply.