Anti-Dengue Virus 1+2+3+4 antibody (ab9200)

I received a new vial of ab31469, but I'm still not seeing any signal in ELISA even at 1:10 dilution.

ANSWER:

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of ab9200.

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

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

I would like to accept your offer of a refund. Please let me know how to proceed.

ANSWER:

Thank you for contacting us.

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I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

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I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

1) Abcam product code ab ab9200

2) Abcam order reference number or product batch number LOT: GR7752-4

3) Description of the problem

Many, many, a-specific bands. The antibody is supposed to recognize only Dengue virus. Despite this, we see a multitude of bands in the uninfected cells lysate. No additional/specific bands are observed in the Dengue type 2 infected cell lysates.

4) Sample preparation:

Type of sample (whole cell lysates, fraction, recombinant protein…):

Whole cell lysate

Lysis buffer

Cell pellet was directly taken up in Laemli buffer: 62.5 mM Tris-Hcl, PH 6.8, 25% glycerol, 2%SDS, 0.01% Bromophenol Blue, 5% B-mercaptoethanol

Protease inhibitors:

Cell pellet was directly taken up in Laemli buffer: 62.5 mM Tris-Hcl, PH 6.8, 25% glycerol, 2%SDS, 0.01% Bromophenol Blue, 5% B-mercaptoethanol

Phosphatase inhibitors

Cell pellet was directly taken up in Laemli buffer: 62.5 mM Tris-Hcl, PH 6.8, 25% glycerol, 2%SDS, 0.01% Bromophenol Blue, 5% B-mercaptoethanol

Reducing agent

5% B-mercaptoethanol

Boiling for ≥5 min? yes/no

First time that I tried, No boiling Second Time that I tried Yes (10 minutes)

Protein loaded ug/lane or cells/lane

30 mg

Positive control

Dengue type 2 infected human cells

Negative control

Uninfected human cells



5) Percentage of gel

4-20% gradient precast

Type of membrane

PVDF

Protein transfer verified

Yes

Blocking agent and concentration

5% milk

Blocking time

2 hours

Blocking temperature

RT



6) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution

1:2000

Diluent buffer

PBS Tween, 5% milk

Incubation time

Overnight

Incubation temperature:

4 degrees



7) Secondary antibody:

Species:

Goat

Reacts against:

Anti-rabbit IgG

Concentration or dilution

1:7500

Diluent buffer

PBS Tween 5% milk

Incubation time

1 hour

Incubation temperature:

RT

Fluorochrome or enzyme conjugate:

HRP



8) Washing after primary and secondary antibodies:

Buffer

PBS Tween

Number of washes

3X 10 minutes each



9)Detection method

ECL

10) How many times have you run this staining?

2X

Do you obtain the same results every time?

Yes, This antibody stains everything under the sun. Notice that it evens stains three proteins in the marker lane!

What steps have you altered to try and optimize the use of this antibody?

First time I did not boil the samples, Second time I did boil the samples, No difference

Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

Document attached Second try with boiled samples. Lane1 Marker, Lane 2 Uninfected cell lysate, Lane 3-10 Dengue type 2 infected cell lysate

ANSWER:

Thank you for your message and for providing this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments and would be pleased to arrange in compensation a refund /credit note or a replacement with an alternative catalog reference : http://www.abcam.com/ab9202or http://www.abcam.com/ab26837 (tested only in ICC/IF).Unfortunately we do not have any alternative lot in stock for the antibody ab9200.

I look forward to hearing from you with details of how you would like to proceed.

We have now spend some time with this antibody in different assays. We always observe that the reactivity of this antibody is below any standards. Uninfected cells stains very brightly with this antibody using IF and no difference is observed when the cells are infected with Dengue. Likewise, uninfected human cell lysates give a multitude of non-specific bands and no discernible band in Dengue infected lysates. We have concluded that this antibody is completely useless in our research.
We hope that you would be willing to replace it with another anti-dengue virus antibody of our choice or a refund.
In all fairness, I have very good experiences with other antibodies sold by Abcam so I was a bit surprised.

ANSWER:

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. 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 have attached only our WBquestionnaire but not the ICC one to speed the process up.


I look forward to receiving your reply.

I would like to clarify something about the purity of antibodies. I am interested in some anti-dengue antibodies, for example in the following link,
(http://www.abcam.com/Dengue-Virus-1-2-3-4-antibody-ab9200.html) and I do not know if when you say that the antibody purity is purified protein G it means that the solution has not
passed through an antigen-specific colunm, which means that it would not be 100% pure. In that case, it has an estimative about the purity percentage? Another thing is about the
Tissue culture supernatant purity, could you compare the purity between these two cases? Protein G purified and Tissue culture supernatant?

ANSWER:

Thank you for your inquiry.


Protein G purification is a way that some of our antibodies are purified from serum, ascites and tissue culture supernatant. It is different that immunogen affinity purification, but both processes result in a purified antibody.


Tissue culture supernatant, on the other hand, is not purified. These antibodies can be expected to have other proteins in them.


Please see this link for more information: http://www.abcam.com/index.html?pageconfig=resource&rid=11270&pid=11287


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