Key Specifications Table
| Key Applications |
|---|
| ELISA, WB, IHC |
| Description | |
|---|---|
| Catalogue Number | 2170 |
| Brand Family | Chemicon® |
| Trade Name |
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| Description | ChemiBLOCKER |
| Overview | The ChemiBLOCKER™ is an unique membrane-blocking agent. As a non-animal protein blocker it offers an alternative to milk-based blocking agents, minimizing the risk of non-specific binding of antibodies to your membrane and lowering your background. ChemiBLOCKER™ may be used at dilutions as high as 20x but for most applications, a 5 to 10 fold dilution is appropriate. ChemiBLOCKER™ is biotin free and will not interfere with ABC detection systems. ChemiBLOCKER™ does contain phosphates and may not be suitable for use with some phospho-specific antibodies. Related Products: • BLOT-QuickBlocker™ A novel blocking protein which produces a semi-clear solution. This allows users the option to examine their blot membranes during the blocking procedure. 200g Catalog Number 2080 • Femto/TBST wash buffer (10X) 250mL Catalog Number 2081 Materials required: • 1X femto/TBST (Catalog Number 2081), 1X TBST, or 1X PBST • primary and secondary antibodies • reagents for immunodetection |
| Product Information | |
|---|---|
| Components |
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| Presentation | A 2X solution of non-animal proteins in 2mM TRIS, pH 8.5, 100mM NaCl, 0.001% Tween-20, and 0.08% Kathon added as a preservative. |
| Applications | |
|---|---|
| Key Applications |
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| Application Notes | Western Blot ELISA PROTOCOL: 1. Make up blocking solution by mixing 1 part ChemiBLOCKER™ with 1 part 1X femto-TBST/PBST (TBST or PBST also work) for blocking PVDF membranes. For nitrocellulose membranes, mix 1 part ChemiBLOCKER™ with 3 parts 1X femto-TBST (TBST or PBST also work). Note:ChemiBLOCKER™ can be used at a wide variety of dilutions, some optimization may be necessary. ChemiBLOCKER™ is NOT recommended for phospho-protein studies as it does contain phosphorylated proteins. 2. Incubate the membrane in blocking solution (10-20 mL) for 60 minutes at room temperature with shaking in a heat-sealable plastic bag. 3. Prepare antibody dilution buffer (ADB). For PVDF membranes mix 1 part ChemiBLOCKER™ with 3 parts 1X femto-TBST/PBST (TBST or PBST also work). Incubate the membrane in 5-10 mL primary antibody, diluted appropriately with ADB, for 90 min at room temperature with shaking. 4. Wash the membrane 3 times in 50 mL of 1X femto-TBST (TBST or PBST also work) for 5 min each time at room temperature with shaking. 5. Incubate the membrane in 5-10 mL secondary antibody, diluted appropriately with ADB, for 90 min at room temperature with shaking. 6. Wash the membrane as in step #4, but add one extra wash (4 washes total). 7. Wash the membrane 3 times quickly (30 sec.) in 50 mL pure deionized water each time and then process the blot, using femto-LUCENT™ detection system (or your favorite system). |
| Dimensions | |
|---|---|
| Volume | 1 L |
| Product Usage Statements | |
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| Usage Statement |
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| Storage and Shipping Information | |
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| Storage Conditions | Store kit components at 2-8°C for up to 1 year from date of purchase. Warm ChemiBLOCKER™ to room temperature and mix thoroughly before dispensing. To avoid possible microbial contamination, dispense all solutions aseptically. Over time, phenolic compounds may precipitate out of solution resulting in the formation of a small amount of dark brown to black particulate material at the bottom of the container. This material will not affect the performance of the blocking compound. |
| Packaging Information | |
|---|---|
| Material Size | 2 X 500 mL |
| Material Package | sufficient for 100 mini gel blots |