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Anti human igm

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Anti-human IgM is a laboratory reagent used for the detection and quantification of human immunoglobulin M (IgM) in biological samples. It is a highly specific antibody that binds to the Fc region of human IgM molecules.

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Lab products found in correlation

Streptavidin bv421, by BioLegend (2 mentions) Anti human cd19, by BioLegend (2 mentions) Anti human cd21, by BioLegend (2 mentions) Anti human cd27, by BioLegend (2 mentions) Histopaque, by Merck Group (2 mentions) Anti cd27 apc, by BioLegend (1 mentions) Facsaria ιιι flow cytometer, by BD (1 mentions) Beckman flow cytometer, by Beckman Coulter (1 mentions) Anti human cd3, by BioLegend (1 mentions) Facsaria, by BD (1 mentions) 40592 v08h2 b, by Sino Biological (1 mentions) Beckman flow cytometry, by Beckman Coulter (1 mentions)

4 protocols using anti human igm

1

Spike-specific Memory B Cell Isolation

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B cells were purified magnetically (STEMCELL Technologies, 17954) and stained with anti-CD27-APC (Biolegend, 356410), anti-human IgM (Biolegend, 314512), anti-human IgG (Biolegend, 410708) and biotinylated spike protein. Spike-specific memory B cells were isolated by flow-cytometric sorting using a BD FACSAria ΙΙΙ flow cytometer (BD Biosciences) from pooled PBMCs. Flow-cytometric data were analyzed with FlowJo.
Dou Y., Xu K., Deng Y.Q., Jia Z., Lan J., Xu X., Zhang G., Cao T., Liu P., Wang X., Wang X., Xu L., Du P., Qin C.F., Liu H., Li Y., Wu G., Wang K, & Lu B. (2023). Development of neutralizing antibodies against SARS-CoV-2, using a high-throughput single-B-cell cloning method. Antibody Therapeutics, 6(2), 76-86.
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2

SARS-CoV-2 Spike Protein Antibody Detection

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The stained peripheral blood mononuclear cells were evaluated in the following procedure, using a Beckman flow cytometer (Beckman Coulter, Inc., California, USA). Antigen probe was obtained, and the biotinylated SARS-CoV-2 Spike RBD protein (40592-V08H2-B, Sino Biological, Beijing, China) was mixed with Streptavidin-BV421 (405225, Biolegend, California, USA) at a 4:1 molar ratio for 1 h. To obtain the peripheral blood mononuclear cells, density gradient centrifugation [Histopaque (10771, Sigma-Aldrich, St Louis, Missouri, USA)], cell cleaning (FACS, With 2% FBS), add antigen probes (1:33.3) and fluorescent-conjugated antibodies [anti-human IgG Fc (410722, Biolegend), anti-human IgM (314524, Biolegend) Antihuman CD3 (300430, Biolegend), anti-human CD19 (302212, Biolegend), anti-human CD21 (354918, Biolegend), anti-human CD27 (356406, Biolegend)] after being mixed and placed at 4°C, 30 min, conduct light-avoidance staining; Test sample (FACS buffer), Analyze the data [FlowJo software (V10.0.7)].
Guo Q., Yang L., Peng R., Gao T., Chu X., Jiang D., Ke D, & Ren H. (2022). Safety and immunogenicity of inactivated COVID-19 vaccine in patients with metabolic syndrome: A cross-sectional observational study. Frontiers in Public Health, 10, 1067342.
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3

Engineering Ramos Cells for BCR Expression

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Ramos cells were engineered to stably express mature B cell receptors (BCR) IgM versions of VRC01, PGT145, and PGT128.by lentiviral transduction of the cells using FEEKW-vectored light chain and IgM heavy chain-expressing lentiviruses. BCR-positive cells were identified by staining with both PE-conjugated mouse anti-human-kappa/lambda (BD Biosciences, San Jose, CA) and anti-human-IgM (Biolegend, San Diego, CA) by flow cytometry and sorted using a FACS Aria (BD Biosciences). Cells were amplified, assessed for BCR expression as before, and resorted for highest-staining cells if needed.
Cale E.M., Gorman J., Radakovich N.A., Crooks E.T., Osawa K., Tong T., Li J., Nagarajan R., Ozorowski G., Ambrozak D.R., Asokan M., Bailer R.T., Bennici A.K., Chen X., Doria-Rose N.A., Druz A., Feng Y., Joyce M.G., Louder M.K., O’Dell S., Oliver C., Pancera M., Connors M., Hope T.J., Kepler T.B., Wyatt R.T., Ward A.B., Georgiev I.S., Kwong P.D., Mascola J.R, & Binley J.M. (2017). Virus-like Particles Identify an HIV V1V2 Apex-Binding Neutralizing Antibody that Lacks a Protruding Loop. Immunity, 46(5), 777-791.e10.
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4

SARS-CoV-2 Spike RBD Protein Binding Assay

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The stained peripheral blood mononuclear cells were tested by Beckman flow cytometry (Beckman Coulter, Inc., California, USA). The specific steps were as follows: first, we mixed the biotinylated SARS-CoV-2 spike RBD protein (40592-V08H2-B, Sino biological, Beijing, China) with streptavidin-BV421 (405225, Biolegend, California, USA) in a 4:1 mole ratio and leave for 1 h to get the antigen probe; second, peripheral blood mononuclear cells were obtained from whole heparinized blood; the density gradient centrifugation was performed by Histopaque (10771, Sigma–Aldrich, St Louis, Missouri, USA) and then cleaned by cytometric staining buffer (FACS, 2%FBS) cells, besides we added antigen probe (1:33.3), fluorescence-coupled antibody [anti-human IgG Fc (410722, Biolegend), anti-human IgM (314524, Anti-human) CD3 (300430, Biolegend), anti-human CD19 (302212, Biolegend), anti-human CD21 (354918, Biolegend), and anti-human CD27 (356406, Biolegend)] into the cells and dyed at 4°C for 30 min under dark condition. After being re-suspended with FACS buffer, the samples were tested on the machine. The data were analyzed by Flow Jo software (V10.0.7).
Yang L., Zeng T., Li Y., Guo Q, & Jiang D. (2024). Poor immune response to inactivated COVID-19 vaccine in patients with hypertension. Frontiers in Medicine, 11, 1329607.
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