Technical Service Guide: Western Blotting

Customer Information

First Name

Last Name

Company/Institution

Lab Position

Phone

Product Information

Catalog Number / Product Name

Lot Number

Purchase Date

Sales Order Number

Problem and Previous Experience

What is the specific problem you are experiencing?

What size bands were expected and what size bands were detected?

Was the blot blank or was a dark background or non-specific bands seen?

Did this same vial of product work in the past? What were the results?

Did other lots of this product work in the past? Which lots? What were the results?

Samples and Controls

From what species (animal) was the sample and what type of sample was used (cell lysate, tissue, purified protein, etc.)?

What method of sample preparation was used?

Was Alomone blocking peptide used as a negative control?

What positive and/or negative controls were used, and what were the results?

SDS-Page

What percentage gel was used?

What type of gel was used (Tris/glycine, Tris/acetate, etc.)?

How much sample was loaded on the gel?

At what temperature were the samples heated before loading (700C, 1000C, other)?

Transfer and Membranes

What type of transfer system was used (wet, semi-dry, other)?

What were the transfer conditions (time, temperature, etc.)?

What type of membrane was used (PVDF or nitrocellulose)?

Blocking

What were the blocking conditions (blocking solution, how long, what temperature. etc.)?

Primary and Secondary Antibodies

How was the primary antibody reconstituted and stored?

At what dilution(s) was the primary antibody used and what was the diluent?

What were the incubation conditions (time, temperature) for the primary antibody?

From which company was the secondary antibody obtained and what type of secondary was being used (HRP or AP conjugated, anti-rabbit, etc.)?

What detection system was used?

At what dilution(s) was the secondary antibody used and what was the diluent?

What were the incubation conditions (time, temperature) for the secondary antibody?

In order to process your request as soon as we can, please attach result images below and make sure to indicate the molecular weight markers:

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If you tried different experiment conditions, please specify conditions you used and provide the corresponding images.

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Technical Service Guide: Western Blotting