Using secondary antibodies, recognizing the heavy and light-chain of the primary antibody for WB detection of IP samples will always result in two bands (the heavy-chain at 50kDa and the light-chain at 25kDa).
These bands are due to the secondary antibody binding to the primary antibody released with the antigen during the immunoprecipitation procedure, or to the recognition of endogenous IgGs from the lysate itself.
Problem:
- Protein of interest is ~50kDa - heavy-chain will obstruct its detection
- Protein of interest is ~25kDa - light-chain will obstruct its detection
Solution:
- VeriBlot for IP secondary antibodies (for IP products <30kDa) - only recognize native (non-reduced) primary antibodies
- Light-chain specific antibodies (for IP products >30kDa) - do not recognize heavy-chains
Download our Veriblot for IP flyer
- Lane 1: 0.5μg of mouse monoclonal
- Lane 2: 0.25μg of mouse monoclonal
- Lane 3: 0.125μg of mouse monoclonal
VeriBlot for IP (for IP products <30kDa and co-immunoprecipitation experiments)
Product name | Conjugate | Product code |
|---|---|---|
HRP | ||
Biotin | ||
HRP | ||
AP | ||
HRP |
Tip: Please make sure to completely reduce the immunoprecipitate and to use milk as a blocking agent.
Light-chain specific secondary antibodies (for IP products >30kDa)
AP conjugated light-chain specific secondary antibodies
| Product name | Applications | Product code |
|---|---|---|
ELISA, Dot Blot, WB, ICC, IHC-Fr | ||
Flow Cyt, ELISA, ICC/IF, WB | ||
Flow Cyt, ICC/IF, ELISA | ||
Flow Cyt, ICC/IF, WB, ELISA | ||
ELISA, WB | ||
ELISA, WB |
HRP conjugated light-chain specific secondary antibodies
| Product name | Applications | Product code |
|---|---|---|
ELISA, Dot Blot, WB, ICC, IHC-Fr | ||
ELISA, ICC/IF, WB | ||
ICC/IF, ELISA | ||
ICC/IF, WB, ELISA | ||
ELISA, WB | ||
ELISA, WB |
View our AbExcel secondary antibodies for exceptional western blots.
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