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Anti igm hrp

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Anti-IgM-HRP is a laboratory reagent that binds to the IgM antibody and is conjugated with horseradish peroxidase (HRP) enzyme. It is commonly used in immunoassays for the detection and quantification of IgM antibodies in biological samples.

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Lab products found in correlation

Anti igg1 hrp, by Southern Biotech (5 mentions) Anti igg2b hrp, by Southern Biotech (5 mentions) Anti igg3 hrp, by Southern Biotech (4 mentions) Goat anti mouse ig, by Southern Biotech (2 mentions) Slow kinetic tmb solution, by Merck Group (2 mentions) Tepc183, by Merck Group (1 mentions) Anti igg2a hrp, by Southern Biotech (1 mentions) Anti tubulin, by Merck Group (1 mentions) Anti rac1, by Merck Group (1 mentions) Anti syk n 19, by Santa Cruz Biotechnology (1 mentions) Anti cav 1 d46g3, by Cell Signaling Technology (1 mentions) Np bsa biotin, by Biosearch Technologies (1 mentions) Anti cd19 6d5, by BioLegend (1 mentions) Anti lyn, by Cell Signaling Technology (1 mentions) Anti cd95 15a7, by Thermo Fisher Scientific (1 mentions) Recombinant mouse baff, by R&D Systems (1 mentions) Tmb substrate, by BioLegend (1 mentions) Streptavidin hrp, by Southern Biotech (1 mentions) Igg2c, by Southern Biotech (1 mentions) Igg2b, by Southern Biotech (1 mentions)

Close spelling variants of this product

Goat anti-mouse IgM-HRP (18 citations) Anti-mouse IgM-HRP (10 citations) HRP-conjugated goat anti-mouse IgM (8 citations) Goat anti-IgM-HRP (5 citations) Mouse Anti-Human IgM-HRP (4 citations) HRP-conjugated anti-human IgG or IgM (4 citations) Goat Anti-Human IgM-HRP (4 citations) Anti-human IgM-HRP (3 citations) HRP-conjugated anti-mouse IgM (3 citations) HRP-conjugated goat anti-mouse IgM or IgG (3 citations)

7 protocols using anti igm hrp

1

Isotype-Specific ELISA Protocol

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Plates were pre-coated with Goat anti-Mouse Ig (Southern Biotech), blocked with PBS-BB (PBS + 0.05% Tween 20 + 1% BSA) and serum was applied. HRP conjugated secondary antibodies anti-IgA-HRP, anti-IgG3-HRP, anti-IgG1-HRP, anti-IgM-HRP, anti-IgG2b-HRP, and anti-IgG-HRP (all from Southern Biotech) were applied and subsequently exposed with a slow kinetic TMB solution (Sigma). The reaction was terminated with 1N HCL and absorbance was measured at 450 nm on a SpectraMax 340 PC plate reader and analyzed using SoftMax Pro 4.8 Software.
Myers D.R., Norlin E., Vercoulen Y, & Roose J.P. (2019). Active Tonic mTORC1 Signals Shape Baseline Translation in Naive T Cells. Cell reports, 27(6), 1858-1874.e6.
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2

Quantitative IgM ELISA Assay

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Plates were coated with anti-IgM (1 μg/mL; Southern Biotech) overnight. Plates were washed with PBS/0.04% Tween-20, PBS, then water before the addition of cell culture supernatant or IgM standard (TEPC183; Sigma-Aldrich) to the appropriate wells. After 4 hours, plates were washed and incubated with anti-IgM-HRP (1 μg/mL; Southern Biotech) for a further 4 hours. Plates were washed and bound IgM was visualized by the addition of 2,2’-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS; Sigma-Aldrich) substrate solution (0.54 mg/mL ABTS, 10.5 mg/mL citric acid, 15 mg/mL trisodium citrate dihydrate, 0.03% hydrogen peroxide).
Trezise S., Kong I.Y., Hawkins E.D., Herold M.J., Willis S.N, & Nutt S.L. (2023). An arrayed CRISPR screen of primary B cells reveals the essential elements of the antibody secretion pathway. Frontiers in Immunology, 14, 1089243.
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3

Quantifying NP-KLH Antibody Responses

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NP-KLH-specific antibodies were measured with 10 μg/ml of NP-BSA (Bioresearch Technologies) as the coating reagent for ELISA. Diluted serum was incubated in the wells for 1 h at room temperature. Then, NP-KLH-specific antibodies (IgG, IgG1, IgG2a, IgG2b and IgM) were detected using goat polyclonal anti-IgG HRP (Southern Biotech), anti-IgG1 HRP (Southern Biotech), anti-IgG2a HRP (Southern Biotech), anti-IgG2bHRP (Southern Biotech), and anti-IgM HRP (Southern Biotech).
Liu L., Hu L., Yang L., Jia S., Du P., Min X., Wu J., Wu H., Long H., Lu Q, & Zhao M. (2021). UHRF1 downregulation promotes T follicular helper cell differentiation by increasing BCL6 expression in SLE. Clinical Epigenetics, 13, 31.
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4

Isotype-Specific ELISA Protocol

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Plates were pre-coated with Goat anti-Mouse Ig (Southern Biotech), blocked with PBS-BB (PBS + 0.05% Tween 20 + 1% BSA) and serum was applied. HRP conjugated secondary antibodies anti-IgA-HRP, anti-IgG3-HRP, anti-IgG1-HRP, anti-IgM-HRP, anti-IgG2b-HRP, and anti-IgG-HRP (all from Southern Biotech) were applied and subsequently exposed with a slow kinetic TMB solution (Sigma). The reaction was terminated with 1N HCL and absorbance was measured at 450 nm on a SpectraMax 340 PC plate reader and analyzed using SoftMax Pro 4.8 Software.
Myers D.R., Norlin E., Vercoulen Y, & Roose J.P. (2019). Active Tonic mTORC1 Signals Shape Baseline Translation in Naive T Cells. Cell reports, 27(6), 1858-1874.e6.
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5

Comprehensive B Cell Immunophenotyping

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The following antibodies were used for immunoblotting: anti-SYK (N-19; Santa Cruz), anti-IgM-HRP (Southern Biotech), anti-Cav1 (D46G3; Cell Signaling), anti-RAC1 (Millipore), anti-LYN (Cell signaling) and anti-Tubulin (Sigma).
The following antibodies were used for flow cytometry: anti-B220-PECy7 (RA3-6B2), anti-IgD (11-26), anti-IgM (polyclonal), anti-CD19 (1D3), anti-CD4 (GK1.5 and RM4-5), anti-CD8 (53-6.7), anti-CD93 (AA4.1), anti-CD69-PECy7 (H1.2F3), anti-CD86-PE (GL-1), anti-GL7, anti-CD95 (15A7) (all from eBioscience), anti-λLC λ1, λ2 & λ3 (R26-46), anti-κLC (187.1), anti-CD45.1 (A20), anti-IgD-FITC (11-26c.2a), anti-CD45.2 (105), Ki-67 (B56) (all from BD Pharmingen), anti-CD19 (6D5) (Biolegend). NP-BSA-biotin was from Biosearch Technologies. Anti-IgM-Fab’2-Alexa 649 was from Jackson ImmunoResearch (goat polyclonal). CSFE was from Sigma.
Minguet S., Kläsener K., Schaffer A.M., Fiala G.J., Osteso-Ibánez T., Raute K., Navarro-Lérida I., Hartl F.A., Seidl M., Reth M, & Del Pozo M.A. (2017). Caveolin-1-dependent BCR nanoscale organization regulates B cell tolerance. Nature immunology, 18(10), 1150-1159.
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6

Antibody and BAFF Quantification by ELISA

Cited 1 time
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ELISA was used to detect antigen-specific IgM, IgG1, IgG2b, and IgG3 titers by capturing serum antibodies using NIP11-OVA (Jackson ImmunoResearch, Inc) coated plates (NP-specific) or methylated BSA and calf thymus DNA (Sigma-Aldrich) coated plates (DNA-specific), as described previously (30 (link), 31 (link)). Bound antibody was detected with anti-IgM-HRP, anti-IgG1-HRP, anti-IgG2b-HRP or anti-IgG3-HRP (Southern Biotech), TMB substrate (Biolegend), and read on a Molecular Devices microplate reader at 450nm. Titer was determined to be lowest serum dilution achieving signal greater than 2x background. BAFF in sera was measured by a sandwich ELISA using BAFF- specific clones 5A8 (capture) and 1C9-biotin (detection) (Axxora) as performed previously (31 (link)). Recombinant mouse BAFF (R&D Systems) served as standard.
Parthasarathy R., Hägglöf T., Hadley J.T., McLennan A., Mattke A., Dudley E.A., Kumagai A., Dong L.Q, & Leadbetter E.A. (2021). Receptor Interacting Protein Kinase Pathways Regulate Innate B Cell Developmental Checkpoints But Not Effector Function in Mice. Frontiers in Immunology, 12, 758407.
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7

Serum Antibody Isotype Analysis in Mice

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Blood for serum antibody analysis was collected from all mice at 49 days of age. Plates were coated with anti-mouse IgM, IgG1, IgG2b, IgG2c, IgG3 or IgA (Southern Biotech, Birmingham, AL, USA) for 24 h before the addition of diluted serum. IgM, IgG1, IgG2b, IgG3 standards were obtained from Sigma-Aldrich, IgG2c standard was from Southern Biotech, and IgA standard was from Organon Tekcika–Cappel (Durham, NC, USA). Serum Ig was detected with anti-IgM-HRP, anti-IgG1-HRP, anti-IgG2b-HRP, anti-IgG2c-HRP, anti-IgG3-HRP, anti-IgA-biotin and streptavidin-HRP (Southern Biotech) and visualised using ABTS substrate (2,2′-Azinobis (3-ethylbenzthiazoline Sulfonic Acid); Sigma-Aldrich). All samples and standards were measured in duplicate.
Trezise S., Karnowski A., Fedele P.L., Mithraprabhu S., Liao Y., D’Costa K., Kueh A.J., Hardy M.P., Owczarek C.M., Herold M.J., Spencer A., Shi W., Willis S.N., Nutt S.L, & Corcoran L.M. (2018). Mining the Plasma Cell Transcriptome for Novel Cell Surface Proteins. International Journal of Molecular Sciences, 19(8), 2161.
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