anti human cd14 efluor 450 plus anti human pstat1 py701 alexa fluor 647  (Cell Signaling Technology Inc)

Journal: Allergy, Asthma & Immunology Research

Article Title: Flow Cytometry for the Diagnosis of Primary Immunodeficiency Diseases: A Single Center Experience

doi: 10.4168/aair.2020.12.2.292

Figure Lengend Snippet: Flow cytometry panels for primary immunodeficiency disease diagnosis

Article Snippet: STAT1 phosphorylation panel , CD3 (SK7), pSTAT1 (pY701) , Becton Dickinson.

Techniques: Flow Cytometry, Phospho-proteomics

Journal: Allergy, Asthma & Immunology Research

Article Title: Flow Cytometry for the Diagnosis of Primary Immunodeficiency Diseases: A Single Center Experience

doi: 10.4168/aair.2020.12.2.292

Figure Lengend Snippet: Flow cytometry and genetic test findings of AR-chronic granulomatous disease patients (n = 2), autosomal dominant hyper-IgE syndrome (n = 1) and STAT1 mutation (n = 1)

Article Snippet: STAT1 phosphorylation panel , CD3 (SK7), pSTAT1 (pY701) , Becton Dickinson.

Techniques: Flow Cytometry, Mutagenesis, Control

Journal: United European Gastroenterology Journal

Article Title: Oesophageal candidiasis and squamous cell cancer in patients with gain-of-function STAT1 gene mutation

doi: 10.1177/2050640616684404

Figure Lengend Snippet: STAT1 gain-of-function in CMC patient 5. (a) STAT1 phosphorylation following stimulation with interferon (IFN) was assessed by flow cytometry. Whole blood from a healthy control and CMC patient 5 was stimulated with 103 IU/ml interferon (IFN)α (IFN-α2b Intron A, Merck Sharp & Dohme) for 30 minutes, cells were fixed, permeablised and stained with Alexa Fluor R 647 STAT1 (pY701) for pSTAT1 and CD3 Pacific Blue antibody for CD3 + T cells (all reagents from BD). Data were collected with a FACSCaliber (BD) and analysed with FlowJo software. Grey = healthy control; black = CMC patient; clear peaks = unstimulated cells; full peaks = IFNα stimulated cells. Median fluorescence intensity (MFI) is shown. (b) qRT-PCR of STAT1-dependent gene expression for CXCL10 and IRF1. In a healthy control and CMC patient 5, peripheral blood mononuclear cells were isolated and cultured in RPMI medium for 4 h with 1 × 103 IU/ml IFNα (IFN-α2b Intron A, Merck Sharp & Dohme), 1 µg/ml IFNγ (R&D Systems). Total RNA was purified using RNeasy Mini Kit (Qiagen) and reverse-transcribed into cDNA. Gene expression analysis for CXCL1o and IRF1 mRNA was performed with custom TaqMan gene expression assays (ABI). Gene expression was normalised to the housekeeping gene 18S rRNA. Data is expressed in the 2 − Ct format, where Ct = Ct (target) – Ct (18S). Results are for n = 3 replicates. Statistical significance was assessed by unpaired, two-tailed t test using Prism software. * = p > 0.05; ** = p > 0.01.

Article Snippet: Whole blood from a healthy control and CMC patient 5 was stimulated with 10 3 IU/ml interferon (IFN)α (IFN-α2b Intron A, Merck Sharp & Dohme) for 30 minutes, cells were fixed, permeablised and stained with Alexa Fluor R 647 STAT1 (pY701) for pSTAT1 and CD3 Pacific Blue antibody for CD3 + T cells (all reagents from BD).

Techniques: Flow Cytometry, Staining, Software, Fluorescence, Quantitative RT-PCR, Expressing, Isolation, Cell Culture, Purification, Two Tailed Test