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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 CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
STAR209PT WB* 50 µl
STAR209P WB* 0.5 ml
STAR209PA WB* 1 ml
TidyBlot Western Blot Detection Reagent:HRP specifically binds to native (non-reduced) antibodies. In contrast to conventional secondary antibodies, TidyBlot therefore enables the detection of immunoblotted target protein bands without interference from denatured IgG. Sources of denatured IgG include endogenous IgGs present in cell/tissue lysate and primary antibodies released from beads during immunoprecipitation experiments. The latter are particularly obstructive when detecting of (co-)immunoprecipitated proteins as IgG heavy (~50 kDa) and light chains (~25 kDa) may mask the protein(s) of interest.

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.

Product Details

Preservative Stabilisers
<0.06% ProClin™ 300

Storage Information

Storage
Store at +4oC. DO NOT FREEZE.
This product should be stored undiluted. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
12 months from date of despatch.

More Information

Acknowledgements
Proclin™ 300 is a trademark of The Dow Chemical Company (“Dow”) or an affiliated company of Dow.
Regulatory
For research purposes only

Applications of TidyBlot

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
Western Blotting 1 1/40 1/400
  1. 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).
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures.
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:

Species

Monoclonal Isotype(s)

Polyclonal

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

Useful Reagents Available

Description Product Code Pack Size Applications List Price Quantity
Rat anti Mouse Kappa Light Chain:HRP MCA152P 0.1 mg E WB
Mouse anti Rabbit Light Chain:HRP MCA6003P 0.5 ml E WB