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Foxp3 staining kit protocol

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The Foxp3 staining kit protocol is a laboratory tool used to detect and analyze Foxp3-expressing cells, which are associated with regulatory T cells. The kit provides a standardized procedure for sample preparation, permeabilization, and intracellular staining of the Foxp3 protein. This tool enables researchers to quantify and characterize Foxp3-positive cell populations within a sample.

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Lab products found in correlation

Hil 17a, by Thermo Fisher Scientific (2 mentions) Live dead fixable yellow or violet dye, by Thermo Fisher Scientific (2 mentions) Lsrfortessa, by BD (2 mentions) Golgistop, by BD (2 mentions)

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Foxp3 staining protocol Foxp3 staining kit

2 protocols using foxp3 staining kit protocol

1

Comprehensive Immune Cell Profiling

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Cells (0.5–2 × 106) were washed with staining buffer (PBS plus 5% FBS) and surface antigens were stained with fluorochrome-conjugated antibodies for 45 min at 4 oC. Antibodies were used at a 1:200 dilution in a final volume of 200 μl. Fluorochrome-conjugated murine (m) and human (h) specific antibodies used were obtained from Biolegend (mCD45, 30-F11; hCD4, OKT4; hCD8α, HIT8a; hCD45, HI30; hIL-17A, BL168; hIFNγ, 4S.B3; CD56, HCD56; hCD45RO, UCHL1; hCCR5, HEK/1/85a; hCXCR4, 12G5; hCD3; OKT3), Life Technologies (hCD3 PE-Texas Red, MCHD0317) and eBiosciences (CD317/tetherin/BST2, 26F8; Ki-67, 20Raj1). All HIV-infected samples were fixed with 4% paraformaldehyde in PBS for virus inactivation. Where indicated, intracellular cytokines were stained using the Foxp3 staining kit protocol (eBiosciences) after cell suspensions were stimulated for 5 h with 50 ng ml−1 phorbol-12-myristate-13-acetate (Calbiochem) and 1 μM ionomycin (Calbiochem) in the presence of 1 × GolgiStop (BD Biosciences) at 37 oC. Dead cells were excluded by staining with Live/Dead fixable Yellow or Violet dye (Life Technologies). Samples were acquired on an LSRII or LSRFortessa (BD Biosciences) instrument. Flow cytometer data were analysed using FlowJo (TreeStar).
Adoro S., Cubillos-Ruiz J.R., Chen X., Deruaz M., Vrbanac V.D., Song M., Park S., Murooka T.T., Dudek T.E., Luster A.D., Tager A.M., Streeck H., Bowman B., Walker B.D., Kwon D.S., Lazarevic V, & Glimcher L.H. (2015). IL-21 induces antiviral microRNA-29 in CD4 T cells to limit HIV-1 infection. Nature Communications, 6, 7562.
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2

Multiparametric Flow Cytometry Profiling

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0.5 to 2 × 106 cells were washed with staining buffer (phosphate buffered saline (PBS, plus 5% FBS) and surface antigens were stained with fluorochrome-conjugated antibodies for 45 minutes at 4 °C. Antibodies were used at a 1:200 dilution in a final volume of 200 μl. Fluorochrome-conjugated murine (m) and human (h) specific antibodies used were obtained from Biolegend (mCD45, 30-F11; hCD4, OKT4; hCD8α, HIT8a; hCD45, HI30; hIL-17A, BL168; hIFNγ, 4S.B3; CD56, HCD56; hCD45RO, UCHL1; hCCR5, HEK/1/85a; hCXCR4, 12G5; hCD3; OKT3), Life Technologies (hCD3 PE-Texas Red, MCHD0317), eBiosciences (CD317/tetherin/BST2, 26F8; Ki-67, 20Raj1). All HIV infected samples were fixed with 4% paraformaldehyde in PBS for virus inactivation. Where indicated, intracellular cytokines were stained using the Foxp3 staining kit protocol (eBiosciences) after cell suspensions were stimulated for 5 hours with 50 ng ml−1 phorbol-12-myristate-13-acetate (PMA, Calbiochem) and 1 μM ionomycin (Calbiochem) in the presence of 1x GolgiStop (BD Biosciences) at 37 °C. Dead cells were excluded by staining with Live/Dead fixable Yellow or Violet dye (Life Technologies). Samples were acquired on an LSRII or LSRFortessa (BD Biosciences) instrument. Flow cytometer data were analyzed using FlowJo (TreeStar).
Adoro S., Cubillos-Ruiz J.R., Chen X., Deruaz M., Vrbanac V.D., Song M., Park S., Murooka T.T., Dudek T.E., Luster A.D., Tager A.M., Streeck H., Bowman B., Walker B.D., Kwon D.S., Lazarevic V, & Glimcher L.H. (2015). IL-21 induces antiviral microRNA-29 in CD4 T cells to limit HIV-1 infection. Nature Communications, 6, 7562.
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