TidyBlot HRP conjugated Western Blot Detection Reagent enables the western blot detection of your protein of interest in immunoprecipitation (IP) samples and tissue lysates without interference from IgG heavy (~50 kDa) and light (~25 kDa) chains. Unlike conventional detection reagents, such as secondary antibodies, TidyBlot Reagent only binds to native antibodies used for the visualization of your target protein.
TidyBlot Reagent is very versatile as it is compatible with a variety of both monoclonal and polyclonal primary antibodies (Figure 1). In addition to convenience, you do not need to purchase multiple dedicated HRP conjugated secondary antibodies when using this reagent.Learn about the Problems TidyBlot Reagent Solves
Fig. 1. Detection of goat, rabbit and sheep IgG polyclonal antibodies with TidyBlot Reagent. HeLa (YAP1 and mTOR) and HEK293 (NFkappaB p65) cell lysates were run on SDS PAGE and transferred onto PVDF membranes. Goat Anti-YAP1 (VPA00104), Rabbit Anti-mTOR (VPA00174) and Sheep Anti-NFkappaB p65 (VPA00015) antibodies were added at a dilution of 1/1000. Detection was performed with TidyBlot Reagent at a 1/200 dilution. All primary antibodies are part of the western blot validated PrecisionAb range.
Please note that TidyBlot Reagent is HRP conjugated and that the western blot bands (red) and protein standards (green) have been pseudocolored.
TidyBlot Reagent detects the IgG monoclonal and polyclonal antibodies listed below:
|Human||IgG1, IgG2, IgG4||Compatible|
IgG2a, IgG2b, IgG3
IgG1: affinity for IgG1 varies and may not be strong
Affinity should therefore be tested on an antibody by antibody basis by performing dot blot analysis.
If using mouse IgG1 monoclonal antibodies, perform a dot blot to determine compatibility. TidyBlot Western Blot Detection Reagent:HRP might not detect mouse IgG1. In that case you could try our HRP conjugated Rat Anti-Mouse Kappa Light Chain Specific Antibody, clone OX-20 (MCA152P). This antibody does not detect mouse IgG heavy chains and thereby allows you to detect immunoprecipitated proteins of ~50 kDa without obstruction by heavy chains. We also offer a comparable product for rabbit; Mouse Anti-Rabbit Light Chain Secondary Antibody, clone SB62a (MCA6003P) is an HRP conjugated secondary antibody that does not bind to rabbit IgG heavy chains.