Journal: STAR Protocols

Article Title: An optimized protocol for phenotyping human granulocytes by mass cytometry

doi: 10.1016/j.xpro.2022.101280

Figure Lengend Snippet:

Article Snippet: CD44 (Clone IM7) - 171Yb , Fluidigm Sciences , Cat#3171003B.

Techniques: Recombinant, Blocking Assay, Electron Microscopy, Red Blood Cell Lysis, Antibody Labeling, Membrane, Software

Journal: STAR Protocols

Article Title: An optimized protocol for phenotyping human granulocytes by mass cytometry

doi: 10.1016/j.xpro.2022.101280

Figure Lengend Snippet: Mass cytometry surface antibody panel

Article Snippet: CD44 (Clone IM7) - 171Yb , Fluidigm Sciences , Cat#3171003B.

Techniques: Mass Cytometry

Journal: STAR Protocols

Article Title: Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes

doi: 10.1016/j.xpro.2021.100537

Figure Lengend Snippet:

Article Snippet: CyTOF: rat anti-mouse CD44-150Nd (IM7) , Fluidigm , Cat#3150018B; RRID: AB_2827882.

Techniques: Recombinant, Staining, Blocking Assay, Saline, Multiplex Assay, Software

Journal: STAR Protocols

Article Title: Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes

doi: 10.1016/j.xpro.2021.100537

Figure Lengend Snippet: CyTOF Antibody Volumes (per 50 μL Maxpar Cell Staining Buffer/sample)

Article Snippet: CyTOF: rat anti-mouse CD44-150Nd (IM7) , Fluidigm , Cat#3150018B; RRID: AB_2827882.

Techniques: Staining

Journal: STAR Protocols

Article Title: Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes

doi: 10.1016/j.xpro.2021.100537

Figure Lengend Snippet: CITE-seq Antibody Volumes (per 50 μL CITE-seq Staining Buffer/sample)

Article Snippet: CyTOF: rat anti-mouse CD44-150Nd (IM7) , Fluidigm , Cat#3150018B; RRID: AB_2827882.

Techniques: Staining

Journal:

Article Title: Mycobacterium bovis BCG-Infected Mice Are More Susceptible to Staphylococcal Enterotoxin B-Mediated Toxic Shock than Uninfected Mice despite Reduced In Vitro Splenocyte Responses to Superantigens †

doi: 10.1128/IAI.70.8.4148-4157.2002

Figure Lengend Snippet: BCG infectious dose correlates with increased expression of the T-cell activation markers CD44, IL-2Rα, and IL-2Rβ. Unfractionated splenocytes (5 × 105 cells/tube) from each type of spleen were stained with FITC-labeled anti-mouse CD44, anti-mouse CD25/IL-2Rα, or anti-mouse CD122/IL-2Rβ and PE-labeled anti-mouse CD4 or anti-mouse CD8α. The percentages of CD4+ CD44hi, IL-2Rαhi, or IL-2Rβhi CD4+ (white bars) or CD8+ cells (black bars) in the spleen were determined by flow cytometry. Means ± standard errors of the means of three experiments are shown. ∗, P < 0.05; ∗∗, P < 0.01. Purified CD4+ T cells from the spleens of BCG HD-infected mice (day 16) were stained with FITC anti-mouse CD44 and were sorted in an EPICS Elite ESP (Beckman Coulter) into CD44low (naïve) and CD44hi (memory/effector) populations. Sorted cells were cocultured at 105 cells/well either with bone marrow-derived dendritic cells (5 × 104 cells/well) in the presence or absence of SEB (10 μg/ml) or with M12 B-lymphoma cells (5 × 104 cells/well) in the presence or absence of BCG-Ag (10 μg/ml). IFN-γ levels were determined as described in Materials and Methods. Mean ± standard deviation of triplicate wells is shown.

Article Snippet: For flow cytometry analysis, splenocytes (5 × 10 5 ) were stained with all or some of the following MAbs (all at 1:100 dilutions in R10A): fluorescein isothiocyanate (FITC)-conjugated mouse anti-mouse Vβ8 TCR (F23.1), FITC-conjugated rat anti-mouse CD25 (IL-2 Rα; 7D4), FITC-conjugated rat anti-mouse CD122 (IL-2 Rβ; TM-β1), FITC-conjugated rat anti-mouse I-A d /I-E d (2G9) (all from PharMingen), or FITC-conjugated rat anti-mouse CD44 (IM7.8.1; Cedarlane Laboratories, Hornby, Canada).

Techniques: Expressing, Activation Assay, Staining, Labeling, Flow Cytometry, Purification, Infection, Derivative Assay, Standard Deviation

Journal: Scientific reports

Article Title: Melanocortin-4 receptor in macrophages attenuated angiotensin II-induced abdominal aortic aneurysm in mice.

doi: 10.1038/s41598-023-46831-4

Figure Lengend Snippet: Figure 4. An osteopontin receptor CD44 mediates vascular vulnerability of MC4RTB/TB mice. (A) Leptin (100 nM)-induced Spp1 gene expression in primary vascular smooth muscle cells from WT male mice pretreated with or without the PI3K inhibitor LY294002 (LY, 10 nM; n = 4). (B) Representative pictures and (C) quantification of osteopontin (OPN) immunostaining in the aorta of WD-fed MC4R+/+ and MC4RTB/TB mice (n = 6, scale bar is 100 μm). (D) Representative pictures of DAPI, α-smooth muscle cells, and merged images in the aorta of WD-fed MC4RTB/TB mice (scale bars represent 20 μm). (E) Changes in the sBP of 14–18-weeks- old MC4RTB/TB mice during Ang II (500 ng/kg/min) infusion concomitantly administered with anti-CD44 or control antibodies (Ab; n = 6–12). (F) Representative pictures of the aorta and (G) AAA incidence and (H) diameter after Ang II infusion into WD-fed MC4R+/+ and MC4RTB/TB mice administered with anti-CD44 or control Abs (n = 6–12). *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: In the experiment of α-MSH (M4135, Sigma-Aldrich, 1 mg/kg/day) or anti-CD44 neutralizing antibody (BE0039, BioXCell, 200 μg/day) treatment, they were injected intraperitoneally into ApoE KO or MC4RTB/TB mice for 4 weeks starting on the day the minipumps were implanted.

Techniques: Gene Expression, Immunostaining, Control

Journal: The American Journal of Pathology

Article Title: Identification and Characterization of Mesenchymal-Epithelial Progenitor-Like Cells in Normal and Injured Rat Liver

doi: 10.1016/j.ajpath.2014.08.029

Figure Lengend Snippet: Primary Antibodies Used in This Study

Article Snippet: All fluorescent secondary antibodies were purchased from Jackson Immunoresearch Laboratories (West Grove, PA) and used at a dilution of 1:200: anti-mouse IgG (DyLight 549), number 715-505-151, subclass 1; anti-mouse IgG (cy2 conjugated, number 115-225-205), subclass 1; anti-mouse IgG (cy3 conjugated, number 115-165-206), subclass 2a; and anti-rabbit IgG (cy2 conjugated, number 715-165-151, and DyLight 488, number 715-095-151). table ft1 table-wrap mode="anchored" t5 caption a7 Antibodies Isotype Company or producer Catalog no. Dilution Mouse anti-KRT18 IgG1 Abcam (Cambridge, MA) Ab668 1:200 Rabbit anti-KRT18 Polyclonal Novus Biologicals, LLC (Littleton, CO) NBP1-67610 1:150 Rabbit anti-human CLDN 7 Polyclonal Abcam Ab27487 1:100 Rabbit anti-GFP Polyclonal Novus Biologicals, LLC NB-600-308 1:1000 Rabbit anti-rat Desmin Polyclonal AnaSpec, Inc. (San Jose, CA) 29593 1:200 Mouse anti-rat THY1/CD90 IgG1 Cedarlane (Hornby, ON, Canada) CL005AP 1:50 Mouse anti-rat THY1/CD90 IgG1 BD Biosciences (San Jose, CA) 553016 1:50 Mouse anti-rat THY1/CD90 IgG2a Antibody Solutions (Sunnyvale, CA) Ab53226 1:25 Mouse anti-CDH1 IgG2a Transduction Lab, BD Biosciences 61081 1:50 Mouse anti-CDH1 IgG1 ECM Biosciences (Versailles, KY) CM1681 1:50 Rabbit anti-GFP Polyclonal Novus Biologicals, LLC AB-4 1:500 Mouse anti-rat CD44 IgG2a Cedarlane CL044 1:50 Mouse OV6 IgG1 Kind gift from Dr. Stewart Sell (Wadsworth Center, Albany, NY) NA 1:50 Mouse anti-ACTA2 IgG2a Abcam Ab11003 1:100 Mouse anti-GFAP IgG1 Abcam Ab80842 1:400 Mouse anti-rat VCAM1 IgG1 Bio Legend (San Diego, CA) 200401 1:100 Rabbit anti-CD29/integrin B1 Monoclonal Epitomics (Burlingame, CA) 2288-1 1:50 Mouse anti-vimentin IgG1 Lab Vision Corporation (Fremont, CA) MS129-PO 1:50 Mouse anti-rat NT5E IgG1 BD Biosciences 551123 1:50 Open in a separate window NA, not applicable.

Techniques: Transduction

Journal: The American Journal of Pathology

Article Title: Identification and Characterization of Mesenchymal-Epithelial Progenitor-Like Cells in Normal and Injured Rat Liver

doi: 10.1016/j.ajpath.2014.08.029

Figure Lengend Snippet: Primers Used in This Study

Article Snippet: All fluorescent secondary antibodies were purchased from Jackson Immunoresearch Laboratories (West Grove, PA) and used at a dilution of 1:200: anti-mouse IgG (DyLight 549), number 715-505-151, subclass 1; anti-mouse IgG (cy2 conjugated, number 115-225-205), subclass 1; anti-mouse IgG (cy3 conjugated, number 115-165-206), subclass 2a; and anti-rabbit IgG (cy2 conjugated, number 715-165-151, and DyLight 488, number 715-095-151). table ft1 table-wrap mode="anchored" t5 caption a7 Antibodies Isotype Company or producer Catalog no. Dilution Mouse anti-KRT18 IgG1 Abcam (Cambridge, MA) Ab668 1:200 Rabbit anti-KRT18 Polyclonal Novus Biologicals, LLC (Littleton, CO) NBP1-67610 1:150 Rabbit anti-human CLDN 7 Polyclonal Abcam Ab27487 1:100 Rabbit anti-GFP Polyclonal Novus Biologicals, LLC NB-600-308 1:1000 Rabbit anti-rat Desmin Polyclonal AnaSpec, Inc. (San Jose, CA) 29593 1:200 Mouse anti-rat THY1/CD90 IgG1 Cedarlane (Hornby, ON, Canada) CL005AP 1:50 Mouse anti-rat THY1/CD90 IgG1 BD Biosciences (San Jose, CA) 553016 1:50 Mouse anti-rat THY1/CD90 IgG2a Antibody Solutions (Sunnyvale, CA) Ab53226 1:25 Mouse anti-CDH1 IgG2a Transduction Lab, BD Biosciences 61081 1:50 Mouse anti-CDH1 IgG1 ECM Biosciences (Versailles, KY) CM1681 1:50 Rabbit anti-GFP Polyclonal Novus Biologicals, LLC AB-4 1:500 Mouse anti-rat CD44 IgG2a Cedarlane CL044 1:50 Mouse OV6 IgG1 Kind gift from Dr. Stewart Sell (Wadsworth Center, Albany, NY) NA 1:50 Mouse anti-ACTA2 IgG2a Abcam Ab11003 1:100 Mouse anti-GFAP IgG1 Abcam Ab80842 1:400 Mouse anti-rat VCAM1 IgG1 Bio Legend (San Diego, CA) 200401 1:100 Rabbit anti-CD29/integrin B1 Monoclonal Epitomics (Burlingame, CA) 2288-1 1:50 Mouse anti-vimentin IgG1 Lab Vision Corporation (Fremont, CA) MS129-PO 1:50 Mouse anti-rat NT5E IgG1 BD Biosciences 551123 1:50 Open in a separate window NA, not applicable.

Techniques: Sequencing, Amplification

Journal: The American Journal of Pathology

Article Title: Identification and Characterization of Mesenchymal-Epithelial Progenitor-Like Cells in Normal and Injured Rat Liver

doi: 10.1016/j.ajpath.2014.08.029

Figure Lengend Snippet: Immunofluorescence microscopy analysis for expression of mesenchymal and epithelial cell marker proteins in 13-11-3-2 cell line. Mesenchymal cell markers were as follows: vascular cell adhesion molecule 1 (VCAM; A); Desmin (B); ACTA2 (α smooth muscle actin; C); glial fibrillary acidic protein (GFAP; F); Vimentin (G); Thy1 (CD90; H); CD73 (Ecto-5′-Nucleotidase; K); and CD29 (Integrin β1; L); Epithelial cell markers were as follows: KRT18 (Cytokeratin 18; D); OV-6 (E); CD44 (I); CDH1 (epithelial cadherin; J); CLDN7 (Claudin-7; M); phase-contrast microphotograph of cultured 13-11-3-5 cells (N); and phase-contrast microphotograph of cultured 13-11-3-3 cells (O). Original magnifications: ×40 (M); ×10 (N and O).

Article Snippet: All fluorescent secondary antibodies were purchased from Jackson Immunoresearch Laboratories (West Grove, PA) and used at a dilution of 1:200: anti-mouse IgG (DyLight 549), number 715-505-151, subclass 1; anti-mouse IgG (cy2 conjugated, number 115-225-205), subclass 1; anti-mouse IgG (cy3 conjugated, number 115-165-206), subclass 2a; and anti-rabbit IgG (cy2 conjugated, number 715-165-151, and DyLight 488, number 715-095-151). table ft1 table-wrap mode="anchored" t5 caption a7 Antibodies Isotype Company or producer Catalog no. Dilution Mouse anti-KRT18 IgG1 Abcam (Cambridge, MA) Ab668 1:200 Rabbit anti-KRT18 Polyclonal Novus Biologicals, LLC (Littleton, CO) NBP1-67610 1:150 Rabbit anti-human CLDN 7 Polyclonal Abcam Ab27487 1:100 Rabbit anti-GFP Polyclonal Novus Biologicals, LLC NB-600-308 1:1000 Rabbit anti-rat Desmin Polyclonal AnaSpec, Inc. (San Jose, CA) 29593 1:200 Mouse anti-rat THY1/CD90 IgG1 Cedarlane (Hornby, ON, Canada) CL005AP 1:50 Mouse anti-rat THY1/CD90 IgG1 BD Biosciences (San Jose, CA) 553016 1:50 Mouse anti-rat THY1/CD90 IgG2a Antibody Solutions (Sunnyvale, CA) Ab53226 1:25 Mouse anti-CDH1 IgG2a Transduction Lab, BD Biosciences 61081 1:50 Mouse anti-CDH1 IgG1 ECM Biosciences (Versailles, KY) CM1681 1:50 Rabbit anti-GFP Polyclonal Novus Biologicals, LLC AB-4 1:500 Mouse anti-rat CD44 IgG2a Cedarlane CL044 1:50 Mouse OV6 IgG1 Kind gift from Dr. Stewart Sell (Wadsworth Center, Albany, NY) NA 1:50 Mouse anti-ACTA2 IgG2a Abcam Ab11003 1:100 Mouse anti-GFAP IgG1 Abcam Ab80842 1:400 Mouse anti-rat VCAM1 IgG1 Bio Legend (San Diego, CA) 200401 1:100 Rabbit anti-CD29/integrin B1 Monoclonal Epitomics (Burlingame, CA) 2288-1 1:50 Mouse anti-vimentin IgG1 Lab Vision Corporation (Fremont, CA) MS129-PO 1:50 Mouse anti-rat NT5E IgG1 BD Biosciences 551123 1:50 Open in a separate window NA, not applicable.

Techniques: Immunofluorescence, Microscopy, Expressing, Marker, Cell Culture

Journal: Cell reports

Article Title: Neutral ceramidase-dependent regulation of macrophage metabolism directs intestinal immune homeostasis and controls enteric infection

doi: 10.1016/j.celrep.2022.110560

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CD44 (162Dy) , Fluidigm , Cat# 3162030B; RRID: AB_2814898.

Techniques: Recombinant, Western Blot, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, SYBR Green Assay, Cell Culture, IP Phosphatase Assay, Software