Western blotting (WB) is regarded as one of the most sensitive techniques to detect specific proteins. The final step of a WB experiment is the visualization of specific proteins, which can be performed in two different ways:
1. Using a primary antibody linked to an enzyme such as alkaline phosphatase (AP) or horseradish peroxidase (HRP)
2. Incubation with a labeled secondary antibody. The advantage of using a secondary antibody is signal enhancement - as several secondary antibodies bind to a single primary.
Our new comprehensive AbExcel secondaries range
comprises 19 AP and HRP-conjugated secondaries for the WB detection of
chicken IgY and goat/mouse/rabbit/rat IgG:
| Conjugate | anti-Chicken IgY | anti-Goat IgG | anti-Mouse IgG | anti-Rabbit IgG | anti-Rat IgG | |||||
|---|---|---|---|---|---|---|---|---|---|---|
| Goat | Rabbit | Donkey | Rabbit | Goat | Rabbit | Donkey | Goat | Goat | Rabbit | |
| AP | - | ab97137 | ab97107 | ab97097 | ab97020 | ab97043 | ab97061 | ab97048 | ab6846 | ab6735 |
| HRP | ab97135 | ab97140 | ab97110 | ab97100 | ab97023 | ab97046 | ab16284 | ab97051 | ab97057 | ab102172 |
Five reasons for using AbExcel secondaries
All lanes: beta actin antibody ab8226 at 1 µg/ml
Lanes 1-2: AbExcel secondary ab97043; 1/5000
Lanes 3-4: AbExcel secondary ab97043; 1/20,000
Lanes 5-6: AbExcel secondary ab97043; 1/50,000
✔ All AbExcel secondaries have been extensively tested
✔ AP-conjugated AbExcel secondaries can be routinely
used at dilutions ranging from 1/5000 to 1/50,000
✔ Based on a 1/5000 dilution in 3ml milk/BSA, you can do over 1500 blots
with a single vial of an AP-conjugated AbExcel secondary
✔ HRP-conjugated AbExcel secondaries can be routinely
used at dilutions ranging from 1/2000 to 1/20,000
✔ Based on a 1/2000 dilution in 3ml milk/BSA, you can do over 600 blots
with a single vial of HRP-conjugated AbExcel secondary
How to improve your WB
Are you having trouble with high background, unequal loading, weak signals or incomplete stripping? Check out our comprehensive WB resources and get some advice on how to improve your experimental conditions:
- Western blot troubleshooting tips
- Stripping for reprobing protocol
- Western blotting - a detailed guide
- Optiblot western blot buffers and accessories
- To accurately determine the protein concentrations of your samples we offer two different types of assay kits utilizing bicinchoninic acid (BCA) (ab102536) or an improved Bradford reagent (ab102535; ab103628)
- Optiblot pre-cast gels - for superior WB experiments; these gels are up to ten times stronger than conventional pre-cast gels and have a one year shelf life
Secondary antibody selector tool
Learn more about pre-adsorbed secondary antibodies
Back to secondary antibody resources
