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TidyBlot

TidyBlot Western Blot Detection Reagent:HRP

Product Type
Accessory Reagent
Specificity
TidyBlot

Product Code Applications Pack Size List Price Your Price Qty
STAR209PT
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
WB * 50 µl loader
List Price Your Price
loader
STAR209PA
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
WB * 1 ml loader
List Price Your Price
loader
STAR209P
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
WB * 0.5 ml loader
List Price Your Price
loader

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.

Pack Size: 50 µl, 1 ml
Store at +4°C. DO NOT FREEZE.
This product should be stored undiluted. Should this product contain a precipitate we recommend microcentrifugation before use.
Pack Size: 0.5 ml
Store at +4°C. DO NOT FREEZE.
This product should be stored undiluted. Should this product contain a precipitate we recommend microcentrifugation before use.

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 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
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
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 loader
List Price Your Price
loader
Description Rat anti Mouse Kappa Light Chain:HRP
Mouse anti Rabbit Light Chain:HRP MCA6003P E WB 0.5 ml loader
List Price Your Price
loader
Description Mouse anti Rabbit Light Chain:HRP

References for TidyBlot

  1. 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.
  2. Kråkenes, T. et al. (2019) CDR2L Is the Major Yo Antibody Target in Paraneoplastic Cerebellar Degeneration.
    Ann Neurol. 86 (2): 316-21.
  3. Wu, J. et al. (2019) Androgen receptor SUMOylation regulates bone mass in male mice.
    Mol Cell Endocrinol. 479: 117-122.
  4. Welkenhuysen, N.et al. (2018) Robustness of Nutrient Signaling Is Maintained by Interconnectivity Between Signal Transduction Pathways.
    Front Physiol. 9: 1964.
  5. Zuliani, I. et al. (2021) The Dysregulation of OGT/OGA Cycle Mediates Tau and APP Neuropathology in Down Syndrome.
    Neurotherapeutics. 18 (1): 340-63.

STAR209PT

152335 159936 160044

STAR209PA

152334 160043 161544

STAR209P

147711 152333 160042 161543

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Please Note: All Products are "FOR RESEARCH PURPOSES ONLY"

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