The iD BlueLadder Revealer is a mouse monoclonal antibody targeting the blue dye of prestained protein weight ladders. In combination with an anti-mouse secondary antibody approprietely conjugated, 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.
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 mouse primary antibody specific to your protein (for dilution, please refer to the instructions from the manufacturer);
- The iD BlueLadder Revealer (mouse mAb; recommended dilution : 1/2000)
Incubate the membrane with an anti-mouse secondary antibody, conjugated.
Perform the detection according to the used conjugate
Note: if HRP is used, 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-HRP instead of the iD BlueLadder Revealer:
- If the primary antibody specific to your protein is HRP-conjugated
- If the primary antibody specific to your protein was not raised in mouse