2170
ChemiBLOCKER
Sinónimos:
Blocking Reagent
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About This Item
Productos recomendados
envase
pkg of 1 L
fabricante / nombre comercial
ChemiBlock
Chemicon®
técnicas
ELISA: suitable
immunohistochemistry: suitable
western blot: suitable
Condiciones de envío
wet ice
Descripción general
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
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
Aplicación
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).
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).
Envase
sufficient for 100 mini gel blots
Componentes
2X ChemiBLOCKER 1 L
Forma física
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.
Almacenamiento y estabilidad
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.
Información legal
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Palabra de señalización
Warning
Frases de peligro
Consejos de prudencia
Clasificaciones de peligro
Aquatic Chronic 2 - Eye Irrit. 2 - Skin Irrit. 2 - Skin Sens. 1
Código de clase de almacenamiento
12 - Non Combustible Liquids
Clase de riesgo para el agua (WGK)
WGK 3
Punto de inflamabilidad (°F)
Not applicable
Punto de inflamabilidad (°C)
Not applicable
Certificados de análisis (COA)
Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»
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