100-105 WBQ

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Technical Support Questionnaire – Western Blot Name: Ching-An Wu Catalog #: NB100-105 Lot Number: AD-1. PO/Order Number: 20120913004 Species, Tissues or Cell Lines Tested: HeLa cell lysate with hypoxic treatment. Test Sample Preparation: Hypoxic treatment was performed by incubating with 0.7% oxygen for 4 hours. Right after the treatment, cells were collected and placed on ice, washed with ice cold PBS, and lysed with ice cold RIPA buffer, and then centrifuged and boiled with sample loading buffer. Antibody Storage Conditions: 4Gel/Electrophoresis Conditions: 8% SDS-PAGE Transfer Conditions: semi-dry Blocking Solution & Duration: 5% milk in TBST. Primary Antibody Diluent and Dilutions Tested: 1:1000 with TBST Primary Antibody Incubation Time and Temperature: 4over night Wash Solution Composition, Repetitions & Times: Click here to enter text. Secondary Antibody Manufacturer, Host Species, Dilution, & Diluent: anti-mouse HRP at 1:5000 with

Transcript of 100-105 WBQ

Page 1: 100-105 WBQ

Technical Support Questionnaire – Western Blot Name: Ching-An Wu Catalog #: NB100-105 Lot Number: AD-1. PO/Order Number: 20120913004

Species, Tissues or Cell Lines Tested: HeLa cell lysate with hypoxic treatment.

Test Sample Preparation: Hypoxic treatment was performed by incubating with 0.7% oxygen for 4 hours. Right after the treatment, cells were collected and placed on ice, washed with ice cold PBS, and lysed with ice cold RIPA buffer, and then centrifuged and boiled with sample loading buffer.

Antibody Storage Conditions: 4℃℃℃℃

Gel/Electrophoresis Conditions: 8% SDS-PAGE

Transfer Conditions: semi-dry

Blocking Solution & Duration: 5% milk in TBST.

Primary Antibody Diluent and Dilutions Tested: 1:1000 with TBST

Primary Antibody Incubation Time and Temperature: 4℃℃℃℃ over night Wash Solution Composition, Repetitions & Times: Click here to enter text.

Secondary Antibody Manufacturer, Host Species, Dilution, & Diluent: anti-mouse HRP at 1:5000 with

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5% milk in TBST

Secondary Antibody Incubation Time & Temperature: Room temp. 1hr Wash Solution Composition, Repetitions, & Times: Click here to enter text.

Detection System, Procedure & Development Time: ECL Molecular weight of band(s): Click here to enter text. Controls: Click here to enter text.

Observations: Incorrect band and too many background band.