Anti-Integrin beta 3 antibody [BV4] (ab7167)

Customer contacted us in search of an integrin beta 3 product which they could conjugate to HRP for use in IHC-P.

ANSWER:

Thank you for contacting us. I have attempted to contact you by phone but thought I might communicate this information by email as well.

I have reviewed of our catalog for mouse monoclonalAnti-Integrin beta 3 antibodies for you. I have also spoken with the lab regarding additives in the buffers of these products which might interfere with conjugation of HRP. After this I can say that AB110131 and AB7167 are the best options. However, each does need to be processed to remove stabilizers and preservatives which will interfere with HRP. At this time we do not offer any other alternatives which do not have these additives.

To remove the preservatives, such as sodium azide, a buffer exchange/purification will need to be performed. OurAntibody Concentration Kit ab102778 (http://www.abcam.com/Antibody-Concentration-Kit-3-columns-ab102778.html)can remove these additives quickly and efficiently. The ‘clean’ antibody may then be conjugated using our EasyLink HPR conjugation kits (http://www.abcam.com/index.html?pageconfig=resource&rid=13148,http://www.abcam.com/EasyLink-HRP-Conjugation-Kit-3-x-10%C2%B5g-HRP-ab102892.html).

We currently, until 31 May 2012, have a promotion whereas you can receive a free EasyLink conjugation kit with purchase of a primary antibody by using the code:

These products are covered by our Abpromise guarantee. We provide scientific support, replacement or refund should this product not perform as indicated on the datasheet. More information on our Abpromise may be found at the following link:

http://www.abcam.com/index.html?pageconfig=abpromise

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.

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

Hi Tech,

My customer has tried the mentioned protocol. It still no showing any staining.

I hope you can send us the replacement as soon as possible as the user is currently need it very urgent for her work.

Thanks

Best regards,

ANSWER:

Thank you for kindly confirming these details. I am sorry this vial of antibody has not worked for this customer.

As requested, we are pleased to arrange a free of charge replacement. I can confirm that 1 vial of ab7167 has been added to your order number OBH041222. This has Abcam order reference 1079357.

I would like to reassure you that this free of charge replacement vial is also covered by our Abpromise guarantee. Should the customer still be experiencing difficulties with the new vial, or if you have any further questions, please do not hesitate to let me know.

Thank you for your help and cooperation with this case. Please do not hesitate to contact me if you need anything further.

Hi tech,

Please find the attached document for all the info. User would like to have it replace (if possible) as soon as possible.

Thanks

Best regards,

ANSWER:

Thank you for taking the time to complete our questionnaire.The details you have kindly provided will provide us with vital information for our monitoring of product quality.

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there few tips to provide on this occasion to help improve the results.

I can recommend it would still bebeneficial to consider trying the following:

1.The datasheet states to aliquot and store at -20oC. As this has been kept at 4oC since January, it may have lost some activity. I am pleased to provide an exception on this occasion. However, we recommend to store products as recommended on the datasheet as soon as possible on arrival. This ensure the antibody remains stable and we can continue to provide our guarantee.

2. I can recommend to try overnight incubation at 4oC. This can often provide more efficient and specific staining.

3 Is the secondary antibody an anti mouse IgG or IgG1?

Alternatively, or if these tips do not work, I am pleased to offer you a free of charge replacement or credit note in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Hi Tech,

My customer has bought the antibody above and have started to run it on few control tissues, tonsil, breast and also appendix.

She has used the protocol as below:

Target retrieval pH 6 with Microwave 10 min.

H2O2 for 5 Min

Primary AB7167 1:50 for 30 min

Secondary HRP for 30 min

DAB for 5 min.

The result is no staining observe. Can you suggest any solution for the above matter?

Thanks

ANSWER:

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

I appreciate the timethe customerhas spent in the laboratory and understandtheir concerns. It is regrettable the results have not been succesful.

I would like to reassure you that this antibody is tested and covered by our guarantee forIHC-P and cow and human samples. 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.

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.

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 questionaire.



Order Details
Antibody code: ab7167

Lot number:

Purchase order number
or preferably Abcam order number:


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, low signal, non-specific satining etc.)

Sample (Species/Tissue/Cell Type/Cell Line etc.)

tonsil, breast, appendix.

Please confirm species, is this human?

Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)



Antigen retrieval (Enzymatic method, Heat mediated technique etc.)

Target retrieval pH 6 with Microwave 10 min.

Which buffer has been used? Have different times and antigen retrieval buffers been tried?


Permeabilization step


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


H2O2 for 5 Min

Has there been any block with BSA or serum? Please provide details.

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



1:50 for 30 min
Please provide information on incubation temperature and wash steps. Has overnight incubation at 4oC been tried? This can sometimes provide more efficient and specific staining.


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

HRP for 30 min
Please provide further detials of the secondary antibody


Detection method

DAB for 5 min

Is the secondary and detection kit working well with other primary antibodies?


Positive and negative controls used (please specify)


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



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

Do you obtain the same results every time?
Yes No


What steps have you altered?


Additional Notes


We would appreciate if you are also able to provide and image which would help us to assess the results

I recently purchased 3 antibodies this past summer including ab7167, ab30394, and ab93943. I am now looking to purchase some positive controls. Preferably, I would like to have the same cell lysate used for a positive control for all 3 antibodies to minimize the number of purchases I would need to make. Based on the product sheets of the 3 products though, it looks like each antibody is using a different cell lysate. I was wondering if there was some cell lysate that I could use for all 3 antibodies as positive controls? Thanks.

ANSWER:

Thank you for contacting Abcam. I looked at the 'The Human Protein Atlas' website to see what the expression of Intergrin beta 1,3 and alpha 5 is in different cell lines and tissues. From what I have found it looks as though a good positive control would be to use HeLa whole cell lysate (ab14656), as all three of your proteins are expressed in this cell line. Please let me know if there is anything else I can help you with.