Ordering Information
- : 2650-S
- : 1 kit
Product Images
Western blot of total protein extracts from human placenta incubated with an antibody against GAPDH and developed using commercially available chemilu...
Chemilucent Plus Western Blot Enhancing Kit (Trial size)
Description:
Chemilucent Plus Western Blot Enhancing Kit (Trial size)
Trade Name:
Chemicon (Millipore)
Qty/Pk:
1 kit
Product Overview:
ChemiLucent™ Plus is a chemiluminescent signal enhancing reagent that can be used independently to boost the signal obtained with all ECL based Western blot detection kits. ChemiLucent™ Plus is derived from a powerful, innovative proprietary technology. Unlike conjugated complex- or chemiluminescent substrate enhancers, ChemiLucent™ Plus works through a novel mechanism by increasing antigen and antibody affinity and accessibility. Due to its unique mechanism, ChemiLucent™ Plus can be used with different antibodies and antigens, such as structural proteins (Actin), enzymes (GAPDH) or membrane proteins (EGFR).It can significantly boost Western blot signals, and is compatable with other enhancing mechanisms to achieve multiplied enhancing effects.
Key Applications:
Western Blotting
Application Notes:
REAGENT PREPARATION:
1X TTBS
Add 6.05 g Tris base (50 mM), 8.76 g sodium chloride (150 mM) to 800 mL of distilled water. Adjust pH to 7.5 with HCl. Adjust to 1 liter with distilled water. Add Tween-20 to 0.1% (v/v). Maybe stored for up to 3 months at room temperature.
Blocking Reagent
Weight 5 g of ChemiLucent Plus Blocking Reagent and dissolve in 100 mL of 1X TTBS solution. Make fresh daily.
PROTOCOL:
1. Remove blot from the transfer apparatus and soak in transfer buffer. Under the hood, pour the ChemiLucent Plus solution into a new container. For a 50 cm2 blotting membrane use 10 mL of ChemiLucent Plus. The solution can be reused up to 4 times without losing its enhancing effect. Remove the membrane from the transfer buffer and drain the remaining buffer on the membrane. For the best results do not let the membrane dry completely. Soak the membrane in the ChemiLucent Plus solution under agitation for 2 minutes then remove the membrane and submerge it in 1X TTBS.
2. Block the membrane with 5% non-fat dry milk in 1X TTBS solution for 30 minutes at room temperature under agitation.
3. Prepare the Antibody Binding Buffer by diluting the supplied 20X stock solution 20 times with ddH2O. Make an appropriate dilution of your primary antibody in the 1X Antibody Binding Buffer. 10 mL of primary antibody solution is suggested for a 50 cm2 membrane. Incubate the blot for one hour at room temperature to overnight at 2-8C.
4. Wash the membrane in 1X TTBS for 5 minutes. Repeat 3-4 times.
5. Incubate the blot with the appropriate HRP-conjugated secondary antibody solution diluted with the Antibody Binding Buffer for one hour at room temperature. 10 mL of secondary antibody solution is recommended for a 50 cm2 membrane.
6. Repeat step 4 to remove unbound HRP-conjugate.
7. Continue using the directions from your ECL kit (for example Chemicon catalog number 2600).
1X TTBS
Add 6.05 g Tris base (50 mM), 8.76 g sodium chloride (150 mM) to 800 mL of distilled water. Adjust pH to 7.5 with HCl. Adjust to 1 liter with distilled water. Add Tween-20 to 0.1% (v/v). Maybe stored for up to 3 months at room temperature.
Blocking Reagent
Weight 5 g of ChemiLucent Plus Blocking Reagent and dissolve in 100 mL of 1X TTBS solution. Make fresh daily.
PROTOCOL:
1. Remove blot from the transfer apparatus and soak in transfer buffer. Under the hood, pour the ChemiLucent Plus solution into a new container. For a 50 cm2 blotting membrane use 10 mL of ChemiLucent Plus. The solution can be reused up to 4 times without losing its enhancing effect. Remove the membrane from the transfer buffer and drain the remaining buffer on the membrane. For the best results do not let the membrane dry completely. Soak the membrane in the ChemiLucent Plus solution under agitation for 2 minutes then remove the membrane and submerge it in 1X TTBS.
2. Block the membrane with 5% non-fat dry milk in 1X TTBS solution for 30 minutes at room temperature under agitation.
3. Prepare the Antibody Binding Buffer by diluting the supplied 20X stock solution 20 times with ddH2O. Make an appropriate dilution of your primary antibody in the 1X Antibody Binding Buffer. 10 mL of primary antibody solution is suggested for a 50 cm2 membrane. Incubate the blot for one hour at room temperature to overnight at 2-8C.
4. Wash the membrane in 1X TTBS for 5 minutes. Repeat 3-4 times.
5. Incubate the blot with the appropriate HRP-conjugated secondary antibody solution diluted with the Antibody Binding Buffer for one hour at room temperature. 10 mL of secondary antibody solution is recommended for a 50 cm2 membrane.
6. Repeat step 4 to remove unbound HRP-conjugate.
7. Continue using the directions from your ECL kit (for example Chemicon catalog number 2600).
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Components:
- ChemiLucent Plus Reagent 50 mL
- Antibody Binding Buffer (20X) 60 mL
- ChemiLucent Plus Blocking Reagent 20 grams
Brand Family:
Chemicon
Storage Conditions:
Store the kit at 2-8°C for up to 12 months after date of receipt.
Packaging:
1,200 cm2
Reagent Type:
Western Blotting Reagents
Product Name:
Chemilucent Plus Western Blot Enhancing Kit (Trial size)
Materials Required but Not Delivered:
- Primary antibody
- Secondary antibody
- Chemiluminescent Substrates (Catalog number 2600)
- Wash buffer (1X TTBS)
- Membrane stripping and re-probing buffer (Catalog number 2500)
- Secondary antibody
- Chemiluminescent Substrates (Catalog number 2600)
- Wash buffer (1X TTBS)
- Membrane stripping and re-probing buffer (Catalog number 2500)