Protein MW Marker, Detection, 9-200kD (Biotin)

Blue Ice

9200

Protein MW Marker detection packs are designed to detect molecular weight ladders on Western Blots when using the horseradish peroxidase (HRP) based Western detection system. The pack contains biotinylated protein ladders and HRP-linked anti-biotin antibody. The molecular weight ladders are a mixture of purified proteins covalently coupled to biotin that resolve to 10 bands that have a size range of 9-200kD. The anti-biotin antibody is used to detect biotinylated protein ladders on Western blots. The packs have been optimized for chemiluminescent Western detection procedures.

Source:
The protein ladder consists of 10 proteins ranging in apparent molecular weight from 9-200kD. The 9kD protein is derived from aprotinin purified from bovine lung. The proteins from 20-50kD are paramyosin fragments. The higher molecular weight proteins are fusions of maltose binding protein (MBP) with paramyosin or paramyosin/lacZ fragments.

Kit Components:
P9106-25A, Prestained Protein Marker, Broad Range: Supplied as a liquid in 65mM Tris-HCl, pH 7.0 , 35mM sodium chloride, 1mM Na2EDTA, 2% SDS, 1mM sodium azide, 40mM dithiothreitol (DTT), 0.01% phenol red, 10% glycerol.
P9106-25B, Biotin (HRP): Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.

Molecular Weight (kD):
200, 140, 100, 80, 60, 50, 40, 30, 20, 9
 
Applications:
Suitable for use in Western Blot.

Recommended Dilution:
P9106-25B, Biotin (HRP): 1:1000. Should only be used to detect the marker, P9106-25A, and not other biotinylated proteins.

Detection of Protein Ladders:
1. Thaw protein ladder on ice. Mix well. Aliquot the desired amount of the protein ladder (10ul for mini-gels and 20ul for full length gels) into a separate tube.
2. Heat the ladder to 95-100°C for 2 minutes.
3. After a quick microcentrifuge spin, load directly onto gel. To ensure uniform mobility, load an equal volume of 1X Reducing SDS Loading Buffer into any unused wells.
 
Note:
1. Perform chemiluminescent detection according to the manufacturer’s instructions except, at the stage of secondary antibody incubation with the Blot. Concurrently add the anti-biotin antibody P9106-25B at a dilution of 1:1000. Incubate for one hour at RT with gentle agitation.
2. Proceed as usual with the detection procedure.

Storage and Stability:
Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for at least 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Companion Products:
B1750-34: Biotin (HRP) Pab Gt x
P9106-75: Prestained Protein Marker, Broad Range