This technical support questionnaire summarizes a Western blot experiment testing the BCAR 3 antibody. Mouse corneal tissue cultured in different media was used as the test sample. The sample was prepared using lysis buffer and each lane contained 80 μg of protein. The primary antibody, BCAR 3, was tested at a 1/500 dilution overnight at 4°C. A single band near 93 kDa was observed, consistent with the expected molecular weight. Actin was used as a control.

1. Technical Support Questionnaire – Western Blot
Name: BCAR 3
Catalog #: NBP1-74622
Lot Number: 710AK
PO/Order Number: 20120706001
Species, Tissues or Cell Lines Tested: B6 mice corneal tissue cultured with different media
Test Sample Preparation: use lysis buffer extra protein on ice, each lane protein CON. 80 g
Antibody Storage Conditions: -20 ℃
Gel/Electrophoresis Conditions: 5% stacking gel, 80 V, 30 min; 10% resolving gel, 120 V
Transfer Conditions: 100 V, 400 mA, 90 min
Blocking Solution & Duration: 10 % Skim milk 1h, RT
Primary Antibody Diluent and Dilutions Tested: 1/500 BCAR 3 in 5 % skim milk
Primary Antibody Incubation Time and Temperature: overnight, 4 ℃
Wash Solution Composition, Repetitions & Times: TBST 10 min/4 times
Secondary Antibody Manufacturer, Host Species, Dilution, & Diluent: anti rabbit , 1/5000

2. Secondary Antibody Incubation Time & Temperature: 1h, RT
Wash Solution Composition, Repetitions, & Times: TBST 10 min/4 times
Detection System, Procedure & Development Time: ECL, 15 min
Molecular weight of band(s): 93 kDa
Controls: Actin
Observations: BCAR 3 have band near 60 – 70 kDa.