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TidyBlot™ gallery image 1

25 ug of HeLa cell lysate spiked with 3.5 ug of mouse IgG was run in lanes 2, 3, 5 and 6 under reducing conditions on SDS PAGE using the Bio-Rad V3 system and transferred to PVDF membrane. Bio-Rad Precision Plus MW marker was run in lanes 1 and 4. Mouse anti actin gamma (VMA00049) was used to detect actin gamma (approximate MW of 46 kDa) in lanes 2 and 5 only. Lanes 3 and 6 were used as controls to detect the background signaling of the secondary detection agent (no primary antibody control).. TidyBlot Western Blot Detection Reagent:HRP was used as the secondary reagent in lanes 2 and 3 and an HRP conjugated anti-mouse IgG heavy and light chain antibody was used as the secondary in lanes 5 and 6. The Blot was visualized with the Bio-Rad Chemidoc™ Touch Imaging System. The TidyBlot Western Blot Detection Reagent:HRP only bound to the native actin gamma antibody whereas the standard anti-Mouse IgG heavy and light recognized both the native and denatured antibodies

TidyBlot™ gallery image 2

HeLa (lane 2) cell lysate was run under reducing conditions on SDS PAGE using the Bio-Rad V3 system and transferred to a PVDF membrane. Bio-Rad Precision Plus molecular weight marker was run in lane 1. Rabbit anti Human GAPDH (AHP1628) was used as the primary antibody at a dilution of 1/1000 in lane 2. TidyBlot was used at 1/200 as the secondary detection reagent. Visualization was carried out using the Bio-Rad ChemiDoc with automatic exposure

TidyBlot™ gallery image 3

Western blot analysis of GAPDH IP samples. IP was performed on Jurkat cell lysates using 10 µg (lanes 1 & 7), 5 µg (lanes 2 & 8), 1 µg (lanes 3 & 9) Mouse anti GAPDH antibody (MCA4739), and 10 µg Mouse IgG1 Negative Control (lanes 4 & 10) (MCA1209).

8.75 µl of each IP was loaded onto an AnykD™ Criterion™ TGX Stain-Free™ gel. 8.75 µl Jurkat whole cell lysate (WCL) was run in lanes 5 & 11 while Precision Plus Protein™ Prestained Standards were run in lane 6.

Rabbit anti GAPDH antibody (VPA00187) was used at 1/1000 in lanes 1-5. HRP conjugated Tidyblot™ (STAR209P) was used at 1/200 and visualized on the Bio-Rad Chemidoc™ Touch Imaging System. Arrow points to GAPDH (molecular weight 37 kDa).

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  • TidyBlot™ Western Blot Detection Reagent:HRP
(Rated 5.0 out of 5 based on 1 customer reviews)
    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
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      STAR209PWB*datasheet pdfdatasheet pdf0.5 ml
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      • 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.

        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.
      • Intended Use
      • Product Form
      • Reconstitution
      • Preparation
      • Preservative Stabilisers
        <0.06% ProClinTM 300
      • Purity
      • Approx. Protein Concentrations
        40 μg/ml
      • Reagents In The Kit
      • Preparing The Antibody
      • Test Principle
      • Buffer Solution
      • 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.
      • Acknowledgements
      • Regulatory
        For research purposes only
      • 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 NameYesNoMin DilutionMax Dilution
        Western Blotting(1)1/401/400
        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. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
      • 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:


        Monoclonal Isotype(s)


        HumanIgG1, IgG2, IgG4Compatible
        MouseIgG2a, 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.
        RabbitTotal IgGCompatible
      • Recommended Protocol
      • ELISA
      • Immunohistology
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Western Blotting
      • Instructions For Use
        When using a 1/200 dilution (25μl in 5ml milk protein), TidyBlot™ Western Blot Detection Reagent:HRP is sufficient to perform approximately 20 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.

      Additional TidyBlot™ Formats

      Formats Applications Sizes available
      TidyBlot™ : HRP WB* 0.5 ml
      • Copyright © 2016 Bio-Rad Antibodies (formerly AbD Serotec)

      Recommended Secondary Antibody

        Recommended Negative Isotype Control

          Useful Reagents

            DescriptionProduct CodePack SizeApplicationsList PriceQuantity
            Rat anti Mouse Kappa Light Chain:HRPMCA152P0.1 mgE, WB
            Mouse anti Rabbit Light Chain:HRPMCA6003P0.5 mlE, WB

            Recommended Positive Controls

              Histology Controls

                  • Product Images

                  • TidyBlot™ thumbnail image 1
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                Write your review

                Great for IPs
                Submitted: 21 Jun 2016
                The main purpose our lab got TidyBlot for was in use for detecting p53 in IPs. As p53 runs very close to the heavy chain, it is always a struggle to detect p53 in a counterblot. TidyBlot was able to detect p53 without any heavy chain problems. I would recommend using it at a very high concentration (as noted in the protocol). I tried getting away with using less (1:1000) to conserve the product, but true to the protocol a much higher concentration was needed. Overall a great product that we will continue to use for our IPs!

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