TidyBlot Western Blot Detection Reagent:HRP
TidyBlot Western Blot detection reagent:HRP exclusively binds to native non-denatured antibodies and does not recognize IgGs present in your immunoprecipitate or lysate. By using TidyBlot you can therefore avoid background from IgG heavy and light chains.
- Product Type
- Accessory Reagent
|Product Code||Applications||Datasheet||MSDS||Pack Size||List Price||Quantity|
N.B. Reactivity of TidyBlot Western Blot Detection Reagent:HRP to mouse IgG1 can be variable. Bio-Rad recommend customers assess compatibility with their IgG1 antibodies via dot blot assay.
- Preservative Stabilisers
- <0.06% ProClin™ 300
- Store at +4oC. DO NOT FREEZE.
This product should be stored undiluted. Should this product contain a precipitate we recommend microcentrifugation before use.
- 12 months from date of despatch.
- Proclin™ 300 is a trademark of The Dow Chemical Company (“Dow”) or an affiliated company of Dow.
- For research purposes only
Applications of TidyBlot
|Application Name||Verified||Min Dilution||Max Dilution|
|Western Blotting 1||1/40||1/400|
- 1 TidyBlot Western Blot Detection Reagent:HRP preferentially binds to the native antibody used during the Western Blot procedure rather than the SDS-denatured/reduced form present in lysates/immunoprecititates. Best results are therefore achieved by denaturing the immunoprecipitate/lysate fully (consider the duration of boiling the sample to reduce IgG disulphide bonds and the type/concentration of reducing agent).
- Technical Advice
- Note: If using mouse IgG1, perform a dot blot to determine compatibilty. TidyBlot Western Blot Detection Reagent:HRP might not detect mouse IgG1.
TidyBlot Western Blot Detection Reagent:HRP detects the IgG polyclonal and monoclonal antibodies listed below:
Bovine IgG2 Compatible Goat IgG2 Compatible Human IgG1, IgG2, IgG4 Compatible Mouse 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.
Compatible Rat IgG2c Compatible Rabbit Total IgG Compatible Sheep IgG2 Compatible
- Instructions For Use
- When using a 1/200 dilution (25 μl in 5 ml milk protein), TidyBlot Western Blot Detection Reagent:HRP is sufficient to perform approximately 20 (STAR209P) or 40 (STAR209PA) mini Western Blots.
1. Dilute primary antibody (see technical advice section) to appropriate concentration in blocking buffer
2. Incubate blot with primary antibody
3. Wash blot
4. Dilute TidyBlot Western Blot Detection Reagent:HRP For best results, optimize dilutions for each experiment
5. Incubate blot with TidyBlot Western Blot Detection Reagent:HRP
6. Wash blot
7. Prepare and add HRP substrate according to manufacturers instructions
8. Develop blot