Hrp conjugated goat anti mouse igg1

Manufactured by Thermo Fisher Scientific

Sourced in United States

The HRP)-conjugated Goat anti-mouse IgG1 is a secondary antibody that binds to the Fc region of mouse IgG1 antibodies. It is conjugated with horseradish peroxidase (HRP), which can be used to detect and quantify the presence of mouse IgG1 in various immunoassays.

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Goat anti-mouse HRP (78 citations) Mouse IgG1 (32 citations) Goat anti-mouse IgG1 (22 citations) HRP-conjugated goat anti-mouse (18 citations) Goat anti-mouse IgG1-HRP (6 citations) HRP-anti-mouse (6 citations) Anti-mouse IgG1-HRP (3 citations) HRP-conjugated anti-mouse IgG1 (2 citations)

5 protocols using hrp conjugated goat anti mouse igg1

Evaluating Viral Plasmid Expression in Fish Cells

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The expression of viral plasmids transfected in fish cells was assessed by Western blotting. Cell lysate prepared and separated by SDS-polyacrylamide gel electrophoresis (PAGE), as previously described [61 ]. Samples were electrophoretically transferred to Immobilon^®-P PVDF membrane (MilliporeSigma) and membranes were blocked with 5% (w/v) BSA/TBST (P-753, Boston Bioproducts) for 1 h at RT. Primary antibodies, anti-Myc monoclonal antibody (Myc, Invitrogen) and anti-β-Actin (Sigma) were diluted in TBST at 1:5000 and incubated overnight at 4 °C. Membranes were incubated with the secondary antibody for 1 h at RT, using horseradish peroxidase (HRP)-conjugated Goat anti-mouse IgG1 (Invitrogen) at 1:10,000 dilution in TBST. Immunoreactive bands were visualized with SuperSignal™ West Pico PLUS chemiluminescent substrate (Thermo Scientific) using Amersham Imager 600 (General Electric). Following exposure with anti-Myc antibodies, membranes were stripped using Restore™ PLUS western blot stripping buffer (Thermo Scientific) for 10 min at RT, thereafter, blocked with 5% (w/v) BSA/TBST for 30 min at RT, and re-probed with the anti β-Actin antibody.

Gorgoglione B., Ringiesn J.L., Pham L.H., Shepherd B.S, & Leaman D.W. (2020). Comparative effects of Novirhabdovirus genes on modulating constitutive transcription and innate antiviral responses, in different teleost host cell types. Virology Journal, 17, 110.

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Serum BDNF Protein Detection

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Western blots were performed as described⁵³. Proteins from 1 µl serum were separated on 4–12% Bis-Tris gradient gels (Invitrogen, NP0321BOX) and transferred to nitrocellulose membranes. After blocking for 1 h in 3% BSA (Sigma-Aldrich, A7906), 3% ECL Prime Blocking reagent (GE Healthcare) in TBS-T, membranes were incubated overnight with anti-BDNF 3C11 antibody (1:2000, Icosagen, 327-100). After washing, 1 h incubation with HRP-conjugated goat anti-mouse IgG1 (1:2000, Invitrogen, PA1-74421) followed, and membranes washed and developed.

Dingsdale H., Nan X., Garay S.M., Mueller A., Sumption L.A., Chacón-Fernández P., Martinez-Garay I., Ghevaert C., Barde Y.A, & John R.M. (2021). The placenta protects the fetal circulation from anxiety-driven elevations in maternal serum levels of brain-derived neurotrophic factor. Translational Psychiatry, 11, 62.

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Quantification of Antigen-Specific Antibodies by ELISA

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ELISA was performed to measure the titers of Ag-specific IgG1 and IgG2a antibodies in serum. Nunc 96-well MaxiSorp plates (Thermo Fischer Scientific) were coated with 100 µl 1 µg/ml unmodified LYS in carbonate buffer (pH 9.6) (SSI Diagnostica, Hillerød, Denmark). The plates were incubated at 4 °C overnight and washed with 0.2% Tween (Merck) in PBS (pH 7.2, Gibco Life Technologies™). The plates were blocked for 1.5 h with 2% BSA in PBS. To give a 10-fold dilution curve, 1% BSA in PBS was added to each well and the serum was added. The plates were incubated at RT for 2 h. After washing the plates, they were incubated for 1 h with either horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG1 (Invitrogen, Carlsbad, CA, USA) diluted 1:20,000 in 1% BSA in PBS or HRP-conjugated rabbit anti-mouse IgG2a (Invitrogen) diluted 1:5,000 in 1% BSA in PBS. The plates were developed with 100 μl/well 3,3′,5,5′-tetramethylbenzidine substrate (Kem-En-Tec Diagnostics, Taastrup, Denmark) for 10 min. The reaction was stopped by adding 0.2 M sulfuric acid (VWR, Radnor, PA, USA), and the plates were analyzed using a TECAN Sunrise™ ELISA reader (Tecan Trading) at OD₄₅₀ nm corrected at OD₆₂₀.

Wørzner K., Hvannastein J., Schmidt S.T., Foged C., Rosenkrands I., Pedersen G.K, & Christensen D. (2021). Adsorption of protein antigen to the cationic liposome adjuvant CAF®01 is required for induction of Th1 and Th17 responses but not for antibody induction. European Journal of Pharmaceutics and Biopharmaceutics, 165, 293-305.

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Quantifying Antibody Responses in Trichuris Infection

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Sera were collected from naïve or Trichuris muris infected mice at 19 days post-infection and stored at −30°C for side-by-side analysis. Total circulating IgE concentration was determined using the Mouse IgE ELISA Set (BD Biosciences) by following the manufacturer’s protocol. Serially diluted sera at 1:5, 1:25, and 1:125 were used for IgE detection. Trichuris antigen-specific IgG1 and IgG2c concentrations were measured via antigen-specific ELISA. Briefly, Immulon 4HBX Extra High Binding plates (Thermo Scientific) were coated with E/S Trichuris antigen in ELISA coating buffer (PBS supplemented with 0.1 M sodium carbonate (Sigma-Aldrich), 0.1M sodium bicarbonate (Sigma-Aldrich), and 1mM sodium Azide (Sigma-Aldrich) at pH 9.6). Sera were serially diluted two-fold from 1:20 to 1:2560 and incubated in the antigen-coated Immulon 4HBX plates. Antigen-specific IgG1 or IgG2c bound to the immobilized Trichuris antigen was then detected with HRP-conjugated goat anti-mouse IgG1 (Thermo Scientific) or HRP-conjugated goat anti-mouse IgG2c (Thermo Scientific). For IgE, the plates were developed with TMB substrate (Thermo Scientific) and KPL TMB Stop Solution (SeraCare), and optical density (OD) was measured at 450 nm. For IgG1 and IgG2c, the plates were developed with 1-Step^™ ABTS Substrate Solution (Thermo Scientific), and OD was measured at 405nm.

Cadavid L.F., Shufflebotham C., Ruiz F.J., Yeager M., Hughes A.L, & Watkins D.I. (1997). Evolutionary instability of the major histocompatibility complex class I loci in New World primates. Proceedings of the National Academy of Sciences of the United States of America, 94(26), 14536-14541.

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Quantifying Antigen-Specific Antibodies by ELISA

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For determination of antigen-specific antibodies, 50 µl of natural Pru p 3 (nPru p 3; purified from peach peel as described previously^{22 (link)}) or PE were coated on microtiter plates (5 and 50 µg/ml, respectively; in coating buffer) overnight at 4 °C. After blocking with 10% FCS in PBS for 2 h at RT, 50 µl of serum or intestinal homogenates were incubated for 2 h at RT. Biotinylated anti-mouse IgE (R35-118; BD Biosciences; 1:1000) antibody was incubated for 1 h at RT, followed by 30 min incubation of HRP-labeled streptavidin. For detection of antigen-specific IgG1 (sIgG1) and sIgG2a, HRP-conjugated goat anti-mouse IgG1 (Thermo Fisher Scientific; 1:2000) or rabbit anti-mouse IgG2a (Thermo Fisher Scientific; 1:2000) were used. Antigen-specific antibodies were detected by addition of TMB-substrate followed by measurement of the absorbance at 450 nm. Detection of total IgE, total IgG, total IgA as well as mMCPT-1 was performed using commercial ELISA kits according to the manufacturer’s instruction (Thermo Fisher Scientific, Darmstadt, Germany).

Steigerwald H., Krause M., Gonzalez-Menendez I., Quintanilla-Martinez L., Vieths S., Scheurer S., Albrecht M, & Blanco-Pérez F. (2023). Peach extract induces systemic and local immune responses in an experimental food allergy model. Scientific Reports, 13, 1892.

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