Anti-TAF1B antibody (ab90549)

Dear tech, A customer had problems from ab90549 .As follows: 客户姓名 黄世娇 单位 北京大学  联系电话 13581693510  货号 ab90549    批号 GR35937-2  order no  910836 1) Abcam product code   ab90549              2) Abcam order reference number or product batch number GR35937-2 3) Storage conditions: -20 Centigrade 4) Description of the problem (high background, no band, wrong band size...) No band  5) Sample preparation: Name of the tissue/cell line: HeLa lysate Sample species:Human Type of sample (whole cell lysates, fraction, recombinant protein…): whole cell lysates Lysis buffer: 20 mM Tris-HCl [pH 8.0], 5 mM MgCl2, 100 mM KCl, 1 mM EDTA, 0.5%NP-40, 1mM PMSF Protease inhibitors: cocktial Phosphatase inhibitors: Reducing agent                   Boiling for ≥5 min? yes/no yes, 95 Centigrade for 5min  Protein loaded ug/lane or cells/lane  20ug Positive control: cell lysate detected by another protein antibody Negative control  IgG IP sample  6) Percentage of gel  10% Type of membrane  NC membrane Protein transfer verified              Yes Blocking agent and concentration     3% milk Blocking time             1hour      Blocking temperature                room temperature  7) Primary antibody (If more than one was used, describe in “additional notes”) : Concentration or dilution   1: 500  Diluent buffer   3% milk Incubation time  over night Incubation temperature: 4 Centigrade 8)  Secondary antibody:      Species:  Reacts against: mouse IgG Concentration or dilution  1:2500 Diluent buffer   3%milk Incubation time 1.5 hours Incubation temperature: 37 Centigrade Fluorochrome or enzyme conjugate: HRP 9) Washing after primary and secondary antibodies: Buffer   TBST Number of washes       3 times 10)Detection method   11) How many times have you run this staining? Three times Do you obtain the same results every time?  yes What steps have you altered to try and optimize the use of this antibody? Primary antibody antibody dilution from 1:1000 to 1:500, incubation temperature from 4 centigrade to room temperature  

ANSWER:

Thank you for your email. Thank you for your enquiry regarding ab90549 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody. Additionally, thank you for supplying an image, as this has been very beneficial in understanding your concerns.

I appreciate the time you have spent on these experiments in the laboratory and it is disappointing the results have not been successful.

In order to help you further on this I would like to ask how you would like ot proceed in this case. I can either offer you a free of charge replacement from another or a different antibody against same target. Could you let me know which option you would like to take?

I will look forward to hearing from you soon.