Mouse Immunology Research

Dendritic Cells: Langerin & DC-SIGN


Mouse Langerin (CD207): WB left: Lysate from C57/Bl6 mouse ear epidermis (left), CD207-transfected (middle) and untransfected 293T (right) were probed with purified eBioRMUL.2 and revealed with HRP anti-rat IgG. Right: Staining of mouse epidermal sheets using RMUL.2.(top) or rat IgG2a (14-4321)(bottom). Courtesy Mark Udey and Chris Nagao.			Mouse DC-SIGN (CD209, CIRE): CIRE-transfected CHO cells (left) and mouse splenocytes (right) were surface stained with PE rat IgG2a isotype control (open histogram) or PE anti-mouse CIRE (5H10) (colored histogram). Splenocytes shown are gated on CD11C+ cells.

Cytolytic Markers: Perforin & Granzyme B mAbs optimized for IC Staining


Mouse Perforin: Staining of fixed and permeabilized C57BL/6 splenocytes (left) or IL-2 stimulated (right) with APC anti-mouse CD49b/Pan-NK (DX-5) and PE anti-mouse perforin eBioOMAK-D. Quadrant lines demarcate isotype control staining.			Mouse Granzyme B: C57BL/6 splenocytes were treated with mouse IL-2 for 3 days and stained with FITC anti-mouse CD8a (53-6.7). Subsequently, cells were fixed and permeabilized and stained with PE Rat IgG2b Isotype Control (left) or PE 16G6 (right). Total cells were used for analysis.

Intracellular staining: IL-1β & IL-17


Mouse IL-1β: Thioglycolate elicited peritoneal exudate cells were stimulated overnight with LPS in the presence of monensin. Cells were surface stained with FITC anti-mouse CD11b, (clone M1/70), then fixed, permeabilized, and stained with Alexa Fluor^® 647 anti-mouse IL-1β.		Mouse IL-17: CD4+CD25- T cells purified from C57BL/6 mouse splenocytes were cultured with (day 10) bone marrow-derived dendritic cells in the presence of anti-CD3, anti-IL-4, anti-IFN-γ, recombinant TGF-β, and recombinant IL-6 for 4 days. Cells were then reactivated with PMA/ Ionomycin in the presence of monensin for 5 hrs. Cells were harvested, fixed, permeabilized, and stained with Alexa Fluor^® 647 anti-mouse IL-17 and PE anti-mouse IFN-γ.

ELISA: IL-17, IL-23

Neutralization: IL-23 p19, IL-17

Stem/Lineage Panel: CD34, Prominin-1, Notch-1, F4/80


Mouse CD34: Two-color surface staining of mouse bone marrow with anti-mouse CD34 (RAM34), FITC (left) and APC (right), and anti-mouse Hematopoietic Lineage Flow Panel Biotin followed by SA-PE. Total viable cells were used for analysis.			Mouse Prominin-1 (CD133, AC133): Two-color surface staining of mouse bone marrow with anti-mouse Prominin I (13A4) FITC and anti-mouse CD11b (M1/70) APC. Total viable cells were used for analysis.


Mouse Notch-1: Staining of C57Bl/6 thymocytes with PE Mouse IgG1, K Iso Cntrl (open histogram) or PE anti-mo/hu Notch-1 (mN1A) (colored histogram). CD4-CD8- double-negative thymocytes were used for analysis.		Mouse F4/80: Staining of C57Bl/6 splenocytes with APC anti-mouse CD11b (M1/70) and PE Rat IgG2a Iso Cntrl (left) or PE anti-mouse F4/80 (BM8) (right). Total viable cells were used for analysis.

Innate Immunity

Mouse Toll-Like Receptors 1-11 for Flow & WB

Human Immunology Research

Stem/Lineage Cell Panel: VE-Cadherin, Notch-1, DC-SIGN, DEC-205


Human VE-Cadherin (CD144): Staining of Human Umbilical Vein Endothelial Cells (HUVEC) with PE Mouse IgG1, K Iso Cntrl (open histogram) or PE anti-human CD144 (VE-Cadherin) (colored histogram). Total viable cells were used for analysis.		Mouse/Human Notch-1: Staining of C57Bl/6 thymocytes with PE Mouse IgG1, K Iso Cntrl (open histogram) or PE anti-mo/hu Notch-1 (mN1A) (colored histogram). CD4-CD8- double-negative thymocytes were used for analysis.


Human DC-SIGN (CD209): Surface staining of human monocyte-derived immature dendritic cells with anti-human CD209 (eB-h209) FITC (left), and PE (right). Appropriate isotype controls were used (open histogram). Total viable cells were used for analysis.			Human DEC-205 (CD205): Staining of normal human peripheral blood lymphocytes (left) and mature dendritic cells (right) with FITC Mouse IgG2b Iso Cntrl (open histogram) or FITC MG38 (colored histogram). Total viable cells were used for analysis.

Cytokine ELISA: IL-23, IL-17

IC Staining & ELISPOT: IL-17, TGF-β, Granzyme B


Human IL-17: Human PBMCs were stimulated with TPA/ Ionomycin in the presence of monensin for 5 hours. The cells were fixed and permeabilized and intracellularly stained with PE anti-human CD4, clone RPA-T4 and Alexa Fluor^® 647 anti-human IL-17. Isotype control on the left and anti-IL-17 on the right. Cells in the lymphocytes gate were used for analysis.			Human IL-17 ELISPOT: Left: Human PBMCs cultured for 24 hrs (no mitogen). Right: Human PBMCs activated with PMA/Ionomycin for 24 hrs.

Human TGF-β


Human Granzyme B IC Staining Reagents & ELISPOT: Left: Human peripheral blood cells were stimulated for 2 days with IL-2. The cells were surface stained with FITC CD56 (MEM188) and stained intracellularly with PE GB11. Quadrants demarcate boundary for isotype controls. Right: Mouse splenocytes were stimulated for 3 days with IL-2. Cells were surface stained with CD49d (DX5) and subsequently stained intracellularly with PE GB11.

Neutralization / Blocking: IL-23 p19 & IL-17

Fc Blocking Reagents: FcγR-Binding Inhibitor


Human FcγR-Binding Inhibitor: U937 cells were stained with FITC mouse IgG2a (left plot) or FITC mouse IgG2b (right plot) isotype controls without (purple shading) or with (blue line) pre-treatment with affinity purified Human FcγR-Binding Inhibitor. The red line represents autofluorescence of U937 cells.

Innate Immunity

Human Toll-Like Receptors 1-9 for Flow & WB

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