The iD BlueLadder Revealer is a mouse monoclonal antibody targeting the blue dye of prestained protein weight ladders. It is conjugated to HRP, and in combination with the appropriate HRP substrate (colorimetric or chemiluminescent), it allows visualizing the marker blue bands throughout the whole process of western blotting from gels to membranes and up to X-Ray films. This makes the positioning of the marker on the blot very easy and highly accurate.
The mAb is conjugated to horse radish peroxidase (HRP).
Instruction for Western Blotting application:
Run an SDS-PAGE gel with a pre-stained molecular weight marker containing blue bands and transfer the proteins to a membrane
Block the membrane using an appropriate blocking solution e.g. iD Blocking Kit (ID-WCBLO1-001; Eurogentec)
Incubate the membrane with a mix of primary antibodies containing:
- A primary antibody specific to your protein, possibly HRP-conjugated (for dilution, please refer to the instructions from the manufacturer);
- The iD BlueLadder Revealer-HRP (recommended dilution : 1/2000)
If the primary Ab specific to your protein is HRP-conjugated
If the primary Ab specific to your protein is not HRP-conjouagated, incubate the membrane with a secondary antibody, HRP conjugated.
Perform the detection by adding an HRP substrate
Note: a colorimetric or chemiluminescent detection can be performed using respectively the iD TMB Substrate Kit (ID-SUTMB1) or iD HRP Substrate Kit (ID-SUHRP1) (Eurogentec).
The iD BlueLadder Revealer and the iD BlueLadder Revealer-HRP are compatible with the iD Western 1H Detection Kits (Eurogentec).
IMPORTANT NOTE: use the iD BlueLadder Revealer instead of the iD BlueLadder Revealer-HRP if the primary Ab specific to your protein was raised in mouse