TidyBlot
TidyBlot Western Blot Detection Reagent:HRP
- Product Type
- Accessory Reagent
- Specificity
- TidyBlot
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. |
- Preservative Stabilisers
- <0.06% ProClin 300
- Regulatory
- STAR209PT, STAR209PA: For research purposes only.
- STAR209P: For research purposes only
- Guarantee
- STAR209PT, STAR209PA: 12 months from date of despatch.
- STAR209P: 12 months from date of despatch
- Acknowledgements
- ProClin is a trademark of The Dow Chemical Company (“Dow”) or an affiliated company of Dow.
This product should be stored undiluted. Should this product contain a precipitate we recommend microcentrifugation before use.
This product should be stored undiluted. Should this product contain a precipitate we recommend microcentrifugation before use.
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:
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
-
STAR209PT
When using a 1/200 dilution (25 μl in 5 ml milk protein), TidyBlot Western Blot Detection Reagent:HRP is sufficient to perform approximately 2 (STAR209PT), 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 -
STAR209PA, STAR209P
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
Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
---|---|---|---|---|---|---|---|
Rat anti Mouse Kappa Light Chain:HRP | MCA152P | E WB | 0.1 mg | Log in | |||
List Price | Your Price | ||||||
Log in | |||||||
Description | Rat anti Mouse Kappa Light Chain:HRP | ||||||
Mouse anti Rabbit Light Chain:HRP | MCA6003P | E WB | 0.5 ml | Log in | |||
List Price | Your Price | ||||||
Log in | |||||||
Description | Mouse anti Rabbit Light Chain:HRP |
References for TidyBlot
-
Inoue, S.et al. (2019) Drebrin-like (Dbnl) Controls Neuronal Migration via Regulating N-Cadherin Expression in the Developing Cerebral Cortex.
J Neurosci. 39 (4): 678-91. -
Kråkenes, T. et al. (2019) CDR2L Is the Major Yo Antibody Target in Paraneoplastic Cerebellar Degeneration.
Ann Neurol. 86 (2): 316-21. -
Wu, J. et al. (2019) Androgen receptor SUMOylation regulates bone mass in male mice.
Mol Cell Endocrinol. 479: 117-122. -
Welkenhuysen, N.et al. (2018) Robustness of Nutrient Signaling Is Maintained by Interconnectivity Between Signal Transduction Pathways.
Front Physiol. 9: 1964. -
Zuliani, I. et al. (2021) The Dysregulation of OGT/OGA Cycle Mediates Tau and APP Neuropathology in Down Syndrome.
Neurotherapeutics. 18 (1): 340-63.
Please Note: All Products are "FOR RESEARCH PURPOSES ONLY"
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