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FAQ - WEST-one™ Western Blot Detection Spray
Questions |
Answers | |||
| Q1 |
Will WEST-one allow for stripping the membrane and reprobing with another Ab? | A1 | To detect another protein after stripping, we recommend a PVDF membrane to minimize loss of protein. Colorimetric/chromogenic detection agents will leave a permanent stain on the membrane that can interfere with subsequent detection of targets with similar molecular weight. WEST-one is chemiluminescent and will not leave a stain. It is also more sensitive than colorimetric reagents. | |
| Q2 |
Should one use the same Ab concentrations as with traditional western blots? | A2 | This depends on experimental conditions. We do recommend that you follow the traditional concentrations for initial set-up. | |
| Q3 |
Does WEST-one work the same with PVDF and supported nitrocellulose? | A3 | Yes. It works the same with PVDF and NC. | |
| Q4 |
Is WEST-one toxic to skin or when inhaled? | A4 | WEST-one is NOT very toxic to skin or when inhaled when used as recommended (point away from face and at blot). If you have any concerns, you may wear a surgical mask and goggles. | |
| Q5 |
I have high background, did I do something wrong? | A5 | Make sure you drain excess liquid from the blot by holding upright with tweezers after only 1 minute. Also we recommend spraying 2 pumps (1 ml) for a 5 cm x 5 cm blot. | |
| Q6 |
Why do I see spotting on my blot after detecting signal? | A6 | Three recommendations: i) When dispensing, hold the spray nozzle about 10 cm – 15 cm from the blot. ii) Make sure the blot is perfectly flat with no bubbles underneath (which tend to cause the reagent to pool and spot). iii) Use more reagent; up to 1 ml (2 squirts) for every 25 cm2 (5 cm x 5 cm section). |