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Phosflow fix 1

Manufactured by BD
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Phosflow FIX I is a laboratory instrument designed for the detection and analysis of phosphoproteins. It provides a standardized method for the fixation and permeabilization of cells, enabling the quantification of intracellular phosphorylation events.

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Lab products found in correlation

Facs analyzer, by BD (2 mentions) Golgistop, by BD (2 mentions) Il 17a, by BioLegend (1 mentions) Ifn γ, by BioLegend (1 mentions) Cd8 pe cy7 clone sk1, by BioLegend (1 mentions)

Close spelling variants of this product

BD Phosflow Fix Buffer I BD Phosflow

2 protocols using phosflow fix 1

1

SARS-CoV-2 Spike Protein Stimulation of PBMCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
PBMC from healthy donors (n = 6) (collected before 2020) and patients with COVID-19 who recovered (n = 22) were thawed and resuspended at 1 × 107 cells/ml in RPMI supplemented with 5% human serum. One-hundred microliters of cell suspension (1 × 106 cells) was transferred to wells of a 96-well U-bottom plate containing purified SARS-CoV-2 spike protein (40 μg/ml) or media alone. After 24 hours in a 37°C/5% CO2 incubator, GolgiStop (BD Biosciences, catalog no. 554724) was added for 6 hours. The cells were harvested, fixed, and permeabilized (BD, Phosflow FIX I, catalog no. 557870; Phosflow Perm III, catalog no. 558050) and stained with antibodies to CD3 (BV650) (clone OKT3, catalog no. 317323), CD4 (allophycocyanina (APC)/Fire750) (clone A161A1, catalog no. 357425), CD8 (phycoerythrin (PE)/Cy7) (clone SK1, catalog no. 344711), CD25 APC-CD25 (clone MA-251, catalog no. 356110), IL-17A (PE Dazzle 594, BL168, catalog no. 512335), and IFN-γ (fluorescein isothiocyanate) (clone 4S.B3, catalog no. 502505) according to the manufacturer’s instructions (BioLegend, San Diego, CA). The cells were analyzed on a FACS analyzer (BD LSRFortessa). Electronic gates were placed on live CD4+ or CD8+ cells using FlowJo software (BD Biosciences, Ashland, OR), and the proportions of CD25+ or IFN-γ+ cells were measured. The percentage positive was determined by comparison to an isotype control antibody.
Pierce C.A., Preston-Hurlburt P., Dai Y., Aschner C.B., Cheshenko N., Galen B., Garforth S.J., Herrera N.G., Jangra R.K., Morano N.C., Orner E., Sy S., Chandran K., Dziura J., Almo S.C., Ring A., Keller M.J., Herold K.C, & Herold B.C. (2020). Immune responses to SARS-CoV-2 infection in hospitalized pediatric and adult patients. Science Translational Medicine, 12(564), eabd5487.
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2

SARS-CoV-2 Spike Protein T-cell Response

Check if the same lab product or an alternative is used in the 5 most similar protocols
PBMC from healthy donors (n=6) (collected before 2020) and patients with COVID-19 who recovered (n=22) were thawed and resuspended at 1×107 cells/ml in RPMI supplemented with 5% human serum. 100μL of cell suspension (1×106 cells) was transferred to wells of a 96-well U-bottom plate containing purified SARS-CoV-2 spike protein (40μg/mL) or media alone. After 24 h in a 37°C/5% CO2 incubator, Golgistop (BD Biosciences, Cat#554724) was added for 6 h. The cells were harvested, fixed and permeabilized (BD, Phosflow FIX I (Cat#557870, Phosflow Perm III Cat#558050) and stained with antibodies to CD3 (BV650) (clone OKT3, Cat#317323, CD4 (APC/Fire750) (clone A161A1, Cat#357425, CD8 (PE/Cy7)(clone SK1, Cat#344711), CD25 APC-CD25 (Clone MA-251, Cat#356110), IL-17A (PE Dazzle 594, BL168, Cat512335), and IFNγ (FITC)(clone 4S.B3, Cat#502505) according to the manufacturer’s instructions (Biolegend, San Diego, CA). The cells were analyzed on a FACS analyzer (BD LSRFortessa). Electronic gates were placed on live CD4+ or CD8+ cells using FlowJo software (BD Biosciences, Ashland, OR) and the proportions of CD25+ or IFN-γ+ cells were measured. The percentage positive was determined by comparison to an isotype control antibody.
Pierce C.A., Preston-Hurlburt P., Dai Y., Aschner C.B., Cheshenko N., Galen B., Garforth S.J., Herrera N.G., Jangra R.K., Morano N.C., Orner E., Sy S., Chandran K., Dziura J., Almo S.C., Ring A., Keller M.J., Herold K.C, & Herold B.C. (2020). Immune responses to SARS-CoV-2 infection in hospitalized pediatric and adult patients. Science Translational Medicine, 12(564), eabd5487.
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