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Alexa fluor 647 conjugated mab against phosphorylated

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Alexa Fluor 647-conjugated monoclonal antibody against phosphorylated proteins. This antibody is labeled with the Alexa Fluor 647 fluorescent dye and can be used to detect and quantify phosphorylated proteins in various applications such as Western blotting, flow cytometry, and immunohistochemistry.

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Lab products found in correlation

Cytofix and phosflow perm wash buffers, by BD (2 mentions) Goat f ab 2 anti human iga g m, by Jackson ImmunoResearch (2 mentions) Alexa fluor 488 conjugated mab against phosphorylated, by BD (2 mentions) Aurora cytometer, by FlowJo (2 mentions)

2 protocols using alexa fluor 647 conjugated mab against phosphorylated

1

Phosphorylation of Syk and PLC-γ2 in BCR-stimulated PBMCs

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PBMCs stained with mAbs against CD19, CD20, CD3, CD27, CD21, IgD, and BV421-conjugated RBD-B.1 were resuspended in RPMI 1640–10% FBS and stimulated with anti-BCR as described previously (Kardava et al., 2018 (link)), with the following modifications. The cells were stimulated at 37°C for 2 min with 10 μg/ml goat F(ab’)2 anti-human IgA/G/M (# 109-006-064 from Jackson ImmunoResearch Laboratories). For the detection of phosphorylated signalling intermediates, cells were fixed and permeabilized using BD Cytofix and Phosflow Perm/Wash buffers (BD Biosciences) and stained with Alexa Fluor 488-conjugated mAb against phosphorylated Syk (p-Y348) and Alexa Fluor 647-conjugated mAb against phosphorylated PLC-2 (p-Y759) (BD Biosciences). The samples were acquired on an Aurora cytometer and analysis was performed by FlowJo V10.
Buckner C.M., Kardava L., Merhebi O.E., Narpala S.R., Serebryannyy L., Lin B.C., Wang W., Zhang X., de Assis F.L., Kelly S.E., Teng I.T., McCormack G.E., Praiss L.H., Seamon C.A., Rai M.A., Kalish H., Kwong P.D., Proschan M.A., McDermott A.B., Fauci A.S., Chun T.W, & Moir S. (2022). Recent SARS-CoV-2 infection abrogates antibody and B-cell responses to booster vaccination. medRxiv.
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2

BCR Stimulation and Signaling Analysis

Cited 1 time
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PBMCs stained with mAbs against CD19, CD20, CD3, CD27, CD21, IgD, and BV421-conjugated RBD-B.1 were resuspended in RPMI 1640–10% FBS and stimulated with anti-BCR as described previously (Kardava et al., 2018 (link)), with the following modifications. The cells were stimulated at 37°C for 2 min with 10 μg/ml goat F(ab’)2 anti-human IgA/G/M (# 109-006-064 from Jackson ImmunoResearch Laboratories). For the detection of phosphorylated signaling intermediates, cells were fixed and permeabilized using BD Cytofix and Phosflow Perm/Wash buffers (BD Biosciences) and stained with Alexa Fluor 488-conjugated mAb against phosphorylated Syk (p-Y348) and Alexa Fluor 647-conjugated mAb against phosphorylated PLC-2 (p-Y759) (BD Biosciences). The samples were acquired on an Aurora cytometer and analysis was performed by FlowJo V10.
Buckner C.M., Kardava L., El Merhebi O., Narpala S.R., Serebryannyy L., Lin B.C., Wang W., Zhang X., Lopes de Assis F., Kelly S.E., Teng I.T., McCormack G.E., Praiss L.H., Seamon C.A., Rai M.A., Kalish H., Kwong P.D., Proschan M.A., McDermott A.B., Fauci A.S., Chun T.W, & Moir S. (2022). Interval between prior SARS-CoV-2 infection and booster vaccination impacts magnitude and quality of antibody and B cell responses. Cell, 185(23), 4333-4346.e14.
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