ImmunoOneStep™ Western
ImmunoOneStep™ Western is a ready-to-use solution that combines blocking and primary/secondary antibody reactions into a single step in Western blotting, enabling exceptionally rapid and efficient blocking and antibody reactions.
Compared with conventional skim milk-based methods, it significantly reduces the time required for blocking and antibody reactions.
Features
- Ready-to-use solution
- Completes blocking and primary/secondary antibody reactions in Western blotting in a single reaction in as little as 1 hour
- Protein-free formulation reduces background
≪ Typical Western Blotting Workflow ≫
Instructions for Use
- (1) After transfer, immerse the membrane in PBS-T/TBS-T buffer for 5 minutes.
- (2) Dilute the primary and secondary antibodies separately with this product in individual tubes.
- (3) Add equal volumes of the primary and secondary antibody solutions to the same container.
- (4) Within 10 minutes, immerse the membrane in the solution from step (3) and incubate with shaking at room temperature for 1–2 hours.
- (5) Wash the membrane 3 times with shaking in PBS-T/TBS-T.
- (6) Perform chemiluminescent/chromogenic detection.
Application Data
EGFP
| Lane No. | Total protein amount (ng) |
|---|---|
| 1 | 1 |
| 2 | 3 |
| 3 | 10 |
- (A)
- Conventional Western blotting
- (B)
- Western blotting using this product
- Sample
- EGFP-overexpressing HEK293 cell lysate
- Gel
- SuperSep™ Ace, 10-20%, 17well (Product No. 198-15041)
- Primary antibody
- Anti Green Fluorescent Protein, Monoclonal Antibody(mFX75) (Product No. 012-22541)
- Secondary antibody
- Anti-Mouse IgG HRP Conjugated
Using this product, EGFP overexpressed in HEK293 cells was successfully detected.
With an exposure time of 3.6 seconds, the detection sensitivity was equivalent to conventional Western blotting with a 1-second exposure.
β-Actin
| Lane No. | Total protein amount (ng) |
|---|---|
| 1 | 1 |
| 2 | 3 |
| 3 | 10 |
- (A)
- Conventional Western blotting
- (B)
- Western blotting using this product
- Sample
- EGFP-overexpressing HEK293 cell lysate
- Gel
- SuperSep™ Ace, 10-20%, 17well (Product No. 198-15041)
- Primary antibody
- Anti β-Actin, Monoclonal Antibody (Product No. 010-27841)
- Secondary antibody
- Anti-Mouse IgG HRP Conjugated
Using this product, endogenous β-actin in HEK293 cells was successfully detected.
The detection sensitivity was higher than that of conventional Western blotting.
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For research use or further manufacturing use only. Not for use in diagnostic procedures.
Product content may differ from the actual image due to minor specification changes etc.
If the revision of product standards and packaging standards has been made, there is a case where the actual product specifications and images are different.
The prices are list prices in Japan.Please contact your local distributor for your retail price in your region.