Anti-ZCCHC6 antibody (ab76901)

Our customer had test following your suggestion and I have transfer the file. Check this please.  

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 a replacement or a credit note in compensation.

Please note that, unfortunately, we cannot offer an alternative catalog reference or another lot of ab76901. If your customer wants a replacement it will be from the same lot.

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

LOT NUMBER GR44511-2 ORDER NUMBER ********** DESCRIPTION OF THE PROBLEM Non-specific band SAMPLE Hela cell, whole cell lysate PRIMARY ANTIBODY Concentration or dilution : 1:100, 1:500 Diluent buffer : 1% Milk in PBS-T Incubation time : 2hr & Overnight Incubation temperature: 2hr at RT, Overnight at 4℃ Wash Buffer : PBST (0.1% Tween20) Number of washes 3 times DETECTION METHOD Chemidoc(Biorad) POSITIVE AND NEGATIVE CONTROLS USED - ANTIBODY STORAGE CONDITIONS -20C SAMPLE PREPARATION Lysis buffer : 200mM KCl, 20mM Tris, 0.2mM EDTA, 0.1% Triton-X100 Protease inhibitors: : Yes(Sigma PI cocktail) Phosphatase inhibitors : No Reducing agent : No Boiling for ≥5 min? yes/no : Yes AMOUNT OF PROTEIN LOADED 50ug/lane ELECTROPHORESIS/GEL CONDITIONS Fig1. 10% Fig2. 8% TRANSFER AND BLOCKING CONDITIONS Type of membrane : PVDF Protein transfer verified : WB same blot with another antibody  Yes Blocking agent and concentration : 5% Milk in PBS-T Blocking time : 1hr, Blocking temperature : RT(20℃) SECONDARY ANTIBODY Pierce 31430 Goat anti-mouse IgG(H+L) Concentration or dilution : 1:5000 Diluent buffer : PBST Incubation time : 40min, 1hr Incubation temperature: RT Fluorochrome or enzyme conjugate: HRP Wash Buffer : PBST (0.1% Tween20) Number of washes 3 times HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Primary concentration, incubation time, incubation temperature ADDITIONAL NOTES -

ANSWER:

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results:

When testing our antibodies, our lab uses 5% BSA as a blocking reagent, so I recommend switching to this instead of milk. Some antibodies give stronger, more specific signals on blots blocked with BSA instead of milk, so doing this may improve the results you are seeing, and reduce the non-specific bands. An example of the above is the western blot obtained with the Abcam GAPDH antibody ab9385 : Click here for the western blot image using ab9385 (or use the following: www.abcam.com/ab9385). Should the suggestions not improve the results, please do let me know.

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

I hope this information is helpful, and I thank you for your cooperation.