Pstat1 y701

Manufactured by BD

The PSTAT1-Y701 is a laboratory equipment product that serves a core function. It is a factual and unbiased description of the product without any interpretation or extrapolation.

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Lab products found in correlation

Human pd l1, by Cell Signaling Technology (1 mentions) Fitc conjugated anti human igg, by Merck Group (1 mentions) Human beta actin, by Cell Signaling Technology (1 mentions) Annexin 5 fitc, by BioLegend (1 mentions) Accutase, by Merck Group (1 mentions) Anti foxp3 mab, by Thermo Fisher Scientific (1 mentions) Pstat5 y694, by BD (1 mentions) Facs canto flow cytometer, by Tree Star (1 mentions) Anti pstat3 y705, by BD (1 mentions) Anti ifnγ xmg1.2, by Thermo Fisher Scientific (1 mentions) Anti il 17a ebio17b7, by Thermo Fisher Scientific (1 mentions) Anti cd29 hmβ1 1, by BioLegend (1 mentions) Anti cd3, by Thermo Fisher Scientific (1 mentions) Dnase 1, by Merck Group (1 mentions) Collagenase d, by Roche (1 mentions)

3 protocols using pstat1 y701

TAM Receptor Expression and Activation

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TAM-overexpressing and vector-expressing MCF-10A cells were detached with accutase (Sigma-Aldrich) and washed with PBS and resuspended in 100μl 1% BSA/PBS. For flow cytometry, anti-human Mertk (R&D), PE-conjugated anti-human Axl (R&D) and PE-conjugated anti-human Tyro3 (R&D) were used according to manufacturer protocol. To evaluate binding of PS-targeting agents to apoptotic cells, 1.0×10⁶ apoptotic or live Jurkat cells in 100 μl DMEM were pre-incubated 10 μg/ml of the PS-targeting 11.31 antibody followed by a second incubation of 0.5 μg FITC-conjugated anti-human IgG (Sigma-Aldrich) or 5μl FITC-Annexin V (Biolegend) for 30min at room-temperature. Immunoblotting was performed by standard protocols with the following antibodies: human Mertk (Cell Signaling), human phospho Mertk (FabGennix), human Axl (Santa Cruz), human phospho Axl (Cell Signaling), human Tyro3 (Cell Signaling), human phospho Tyro3 (Aviva Systems), human Akt (Cell Signaling), human phospho Akt (Cell Signaling), pSTAT1-Y701 (BD Bioscience), human PD-L1 (Cell Signaling), human beta-actin (Cell Signaling)

Kasikara C., Kumar S., Kimani S., Tsou W.I., Geng K., Davra V., Sriram G., Devoe C., Nguyen K.Q., Antes A., Krantz A., Rymarczyk G., Wilczynski A., Empig C., Freimark B., Gray M., Schlunegger K., Hutchins J., Kotenko S.V, & Birge R.B. (2017). Phosphatidylserine Sensing by TAM Receptors Regulates AKT-dependent Chemoresistance and PD-L1 Expression. Molecular cancer research : MCR, 15(6), 753-764.

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Multiparametric Flow Cytometry Panel

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MSD^® 96-well Multi-Spot^® tissue culture kit was purchased from Meso Scale Discovery (MSD). Flourochrome-conjugated monoclonal antibodies (mAbs) against CD3, CD4, CD8, CD20, STAT1, pSTAT1-Y701, pSTAT5-Y694, Ki67, and IFN-γ were purchased from BD Biosciences (San Jose, CA). Anti-CD56 mAb (clone: N901) was purchased from Beckman Coulter (Pittsburg, PA). Anti-Foxp3 mAb and IFN-γ neutralizing mAb (clone: NIB42) were purchased from eBioscience (La Jolla, CA).

Sim G.C., Wu S., Jin L., Hwu P, & Radvanyi L.G. (2016). Defective STAT1 activation associated with impaired IFN-γ production in NK and T lymphocytes from metastatic melanoma patients treated with IL-2. Oncotarget, 7(24), 36074-36091.

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Single-Cell Analysis of Immune Cells

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At scarify, single-cell suspensions were isolated from dLNs, spleens, and CNS. Briefly, the brain and spinal cord were obtained, homogenized, and then incubated with collagenase D (2.5 mg/mL, Roche Diagnostics) and DNase I (1 mg/mL, MilliporeSigma) for 30 minutes. Mononuclear cells were enriched by gradient centrifugation at 670g for 30 minutes on a 37%/70% Percoll gradient without interruption. Before staining, cells were blocked with anti-CD16/CD32 antibodies. The following antibodies were used for flow cytometry: anti-CD3 (OKT3), anti-CD45 (30-F11), anti-MHCII (M5/114.15.2), anti-CD8 (53-6.7), anti-CD44 (IM7), anti-Foxp3(FJK-16S), anti–GM-CSF (MP1-22E9), anti-CD49d (R1-2), anti–IL-17A (eBio17B7), and anti–IFN-γ (XMG1.2) (eBioscience); anti-CD4 (GK1.5), anti-CD25 (PC61.5), anti-CD62L (MEL-14), anti–Ly-6G/Gr-1 (1A8-Ly6g),anti-F4/80 (BM8), anti-CCR2 (SA203G11), anti-CCR6 (29-2L17), and anti-CD29 (HMβ1-1) (BioLegend). In addition, anti–p-STAT3 (Y705) (catalog 557814) and p-STAT1 (Y701) (catalog 502069) antibodies were purchased from BD Biosciences. Phosphoflow cytometry analysis was performed using BD Phosflow buffers (554655 and 558050). Stained cells were analyzed by FACSCanto flow cytometer and were analyzed with FlowJo software (Tree Star).

Cao L., Deng J., Chen W., He M., Zhao N., Huang H., Ling L., Li Q., Zhu X, & Wang L. (2023). CTRP4/interleukin-6 receptor signaling ameliorates autoimmune encephalomyelitis by suppressing Th17 cell differentiation. The Journal of Clinical Investigation, 134(4), e168384.

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