Anti-FGFR3 antibody (ab10649)

BATCH NUMBER 421125 ORDER NUMBER -- NOT SPECIFIED --

DESCRIPTION OF THE PROBLEM Week staining

SAMPLE Paraffin-embedded formalin fixed human tissue (colon)

PRIMARY ANTIBODY . DETECTION METHOD . POSITIVE AND NEGATIVE CONTROLS USED . ANTIBODY STORAGE CONDITIONS -20

FIXATION OF SAMPLE formalin

ANTIGEN RETRIEVAL . PERMEABILIZATION STEP

BLOCKING CONDITIONS

SECONDARY ANTIBODY

ADDITIONAL NOTES You recommend this antibody for IHC-P and I would like to know what protocol you have used when you tested it for this application.

ANSWER:

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 can recommend the following procedure, which has been used to test this antibody in IHC-P. This protocol should be a very good starting point, however you may find it requires some optimization in your laboratory.

Tissue Human umbilical cord Fixation Formalin fixed paraffin embedded Enzyme 0.1% TRYPSIN Enzyme incubation 15 - 20 min Enzyme incubation temp. 37°C Special treatment None H2O2 treatment 3%H2O2 in H2O Blocking 5% normal goat serum +1%BSA Blocking time 25 - 35 min Blocking temp. 37°C Tested sample incubation 55 - 65 min 2nd antibody Biotin anti Rabbit IgG (Gt) Biotin APA Rabbit IgG ads/HIgG(gt)

2nd antibody incubation 25 - 35 min ExtrAvidom -HRP ExtrAvidin Kit ExtrAvidin incubation 25 - 35 min Substrate AEC Substrate incubation 5 - 30 min Hematoxylin Yes

Working dilution 1/250 - 1/500 Staining of Amniotic epithelium Tubules Squamous epitelium

I hope it will be helpful to you. Should you continue to have difficulty obtaining results, please do not hesitate to contact me again with a fully completed form. I would be pleased to investigate and offer some further assistance.

Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a credit note/refund will be offered.

Good luck with the next experiments. I look forward to hearing how you get on.

I am testing some antibodies that I purchased from you. Namely, ab10646; ab10647; ab10649; ab5481, in western blot.

However, the results showed many bands and it's hard to identify the correct one.

Other venders sell "blocking peptide" along with a specific antibody to address this issue.

Do you sell "blocking peptide" for the above antibodies?

Thanks

ANSWER:

Thank you for your enquiry.

Further to correspondence with the source of this antibody unfortunately I have been unable to determine whether the multiple bands that you have been observing is typical of any of these antibodies.

As a general rule if a customer has high background on their blot we recommend that they try changing the blocking buffer to BSA, increasing the stringency and duration of the wash conditions and decreasing the concentration of the antibody.

Should you continue to obtain these extraneous bands I would greatly appreciate it if you complete our technical questionnaire by clicking on the links below. This will greatly help our technical team determine the steps that you have taken to optimise this antibody. It would be most useful if you could additionally provide us with the LOT numbers and representative images of the blots.

I look forward to hearing from you.

ab5481

http://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=5481&mode=questionaire

ab10646

http://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=10646&mode=questionaire

ab10647

http://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=10647&mode=questionaire

ab10649

http://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=10649&mode=questionaire