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Cd44 pe im7

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CD44-PE (IM7) is a fluorescently-labeled antibody that binds to the CD44 cell surface antigen. CD44 is a cell-cell and cell-matrix adhesion molecule involved in a variety of cellular functions. This product is suitable for use in flow cytometry applications.

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Lab products found in correlation

Ly6c pe hk1.4, by BioLegend (3 mentions) Cd45 bv510, by BioLegend (2 mentions) Epcam fitc 9c4, by BioLegend (2 mentions) Tryple, by Thermo Fisher Scientific (2 mentions) Aria 2, by BD (2 mentions) Epcam apc cy7, by BioLegend (2 mentions) Nk1.1 bv605, by BioLegend (2 mentions) Aria3 fusion, by BD (2 mentions) Cd90 pe dazzle thy1, by BioLegend (2 mentions) Facscanto 2, by BD (2 mentions) Cd11b pe cy7, by BioLegend (2 mentions) Lsrfortessa, by BD (2 mentions) Cd45 bv510 30 f11, by BioLegend (2 mentions) Cd62l fitc mel 14, by BioLegend (2 mentions) Fc block 2.4g2, by BD (2 mentions) Cd3 apc 17a2, by BioLegend (2 mentions) B220 apc cy7 ra3 6b2, by BioLegend (2 mentions) Cd4 bv421 rm4 5, by BioLegend (2 mentions) Cd8a percp cy5.5 53 6 7, by BioLegend (2 mentions) Ly6g apc 1a8, by BioLegend (2 mentions)

Close spelling variants of this product

CD44-PE (36 citations) CD44 (IM7, (29 citations) PE/CY7 anti-CD44 (IM7, (4 citations) Anti-CD44-PE (IM7, (4 citations) CD44-PE-Cy7 (IM7, (3 citations) Anti-CD44-PE (clone IM7; (2 citations) Anti-CD44-PE-Cy7 (clone IM7, (2 citations)

7 protocols using cd44 pe im7

1

Isolation of Murine and Human Intestinal Cells

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After 4 days of murine and 7 days of human co-culture, Matrigel was disrupted and cells were collected into 15ml falcon tubes. For murine ILC1 cultures Matrigel disruption was not required, and cells were gently rinsed from the bottom of the plate using PBS+2%FCS. Samples were rinsed with PBS, then dissociated in TrypLe (Gibco) for 20mins at 37°C. The sorting buffer after this step contained DNAse (250µg/ml), EDTA (1μl/ml), and HEPES (1μl/ml) to maintain single epithelial cells and avoid clumping. Cells were titruated gently, centrifuged and resuspended in sorting buffer. Cells were then filtered (70µm), having pre-coated the filter with sorting buffer to minimize cell loss, and either rinsed with PBS for fixable Live/Dead staining (UV or nearInfraRed, Thermofisher), or stained with EpCAM, CD45, and the requisite combination of antibodies for the experiment, and analyzed (BD Fortessa) or sorted (BD ARIA3 Fusion & BD Aria 2 using BD FACS Diva 8.0.1 software). Isolation of murine IEC and ILC1 following co-culture was performed using EpCAM–APC Cy7 (G8.8, BioLegend), CD45-BV510 (30-F11, bioLegend), NK1.1 BV605 (PK136, BioLegend), CD44-PE (IM7, BioLegend). Isolation of human IEC, FB, and hILC1 used fixable Live/Dead-UV or Live/Dead-nIR CD45-eFluor450(HI30) Invitrogen, EpCAM-FITC (9C4; BioLegend), CD90-PE/Dazzle (Thy1; BioLegend)
Jowett G.M., Norman M.D., Yu T.T., Arévalo P.R., Hoogland D., Lust S.T., Read E., Hamrud E., Walters N.J., Niazi U., Chung M.W., Marciano D., Omer O.S., Zabinski T., Danovi D., Lord G.M., Hilborn J., Evans N.D., Dreiss C.A., Bozec L., Oommen O.P., Lorenz C.D., da Silva R.M., Neves J.F, & Gentleman E. (2020). ILC1 drive intestinal epithelial and matrix remodelling. Nature materials, 20(2), 250-259.
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2

Isolation of Murine and Human Intestinal Cells

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After 4 days of murine and 7 days of human co-culture, Matrigel was disrupted and cells were collected into 15ml falcon tubes. For murine ILC1 cultures Matrigel disruption was not required, and cells were gently rinsed from the bottom of the plate using PBS+2%FCS. Samples were rinsed with PBS, then dissociated in TrypLe (Gibco) for 20mins at 37°C. The sorting buffer after this step contained DNAse (250µg/ml), EDTA (1μl/ml), and HEPES (1μl/ml) to maintain single epithelial cells and avoid clumping. Cells were titruated gently, centrifuged and resuspended in sorting buffer. Cells were then filtered (70µm), having pre-coated the filter with sorting buffer to minimize cell loss, and either rinsed with PBS for fixable Live/Dead staining (UV or nearInfraRed, Thermofisher), or stained with EpCAM, CD45, and the requisite combination of antibodies for the experiment, and analyzed (BD Fortessa) or sorted (BD ARIA3 Fusion & BD Aria 2 using BD FACS Diva 8.0.1 software). Isolation of murine IEC and ILC1 following co-culture was performed using EpCAM–APC Cy7 (G8.8, BioLegend), CD45-BV510 (30-F11, bioLegend), NK1.1 BV605 (PK136, BioLegend), CD44-PE (IM7, BioLegend). Isolation of human IEC, FB, and hILC1 used fixable Live/Dead-UV or Live/Dead-nIR CD45-eFluor450(HI30) Invitrogen, EpCAM-FITC (9C4; BioLegend), CD90-PE/Dazzle (Thy1; BioLegend)
Jowett G.M., Norman M.D., Yu T.T., Arévalo P.R., Hoogland D., Lust S.T., Read E., Hamrud E., Walters N.J., Niazi U., Chung M.W., Marciano D., Omer O.S., Zabinski T., Danovi D., Lord G.M., Hilborn J., Evans N.D., Dreiss C.A., Bozec L., Oommen O.P., Lorenz C.D., da Silva R.M., Neves J.F, & Gentleman E. (2020). ILC1 drive intestinal epithelial and matrix remodelling. Nature materials, 20(2), 250-259.
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3

Multiparameter Flow Cytometry for Immune Cell Profiling

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Single-cell suspensions in FACs buffer were incubated in Fc block (2.4G2, BD Biosciences, Franklin Lakes, NJ, USA) for 15 min then samples were washed in FACs buffer and centrifuged at 400 × g for 4 min. Cells were then incubated in a solution containing fluorescently labelled antibodies for 30 min in the dark on ice. Following staining, flow cytometry was performed on a BD FACS Canto II (BD Biosciences) or a LSRFortessa™ (BD Biosciences). Analysis was performed on FlowJo software (BD Biosciences) using the gating strategy outlined in Additional file 2: Fig S2.
The following antibodies were used for the detection and phenotyping of immune cells: CD45-BV510 (30-F11; Biolegend, San Diego, CA, USA), CD3-APC (17A2; Biolegend), B220-APC-Cy7 (RA3-6B2; Biolegend), CD4-BV421 (RM4-5; Biolegend), CD8a-PerCP-Cy5.5 (53–6.7; Biolegend), CD62L-FITC (MEL-14; Biolegend), CD44-PE (IM7; Biolegend), CD11b-PE-Cy7 (M1/70; Biolegend), Ly6G-APC (1A8; Biolegend), Ly6C-PE (HK1.4; Biolegend), and CD49d-AF488 (R1-2; Biolegend).
Peck T., Davis C., Lenihan-Geels G., Griffiths M., Spijkers-Shaw S., Zubkova O.V, & La Flamme A.C. (2023). The novel HS-mimetic, Tet-29, regulates immune cell trafficking across barriers of the CNS during inflammation. Journal of Neuroinflammation, 20, 251.
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4

Flow Cytometry for Immune Cell Analysis

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For flow cytometry, the following antibodies were purchased from BD Biosciences: CD25-APC (PC61), CD3-FITC (17A2), CD3-APC-Cy7, CD4-PE (L3T4), CD8a-FITC (LY-2, 53-6.7), CD8a-APC (53-6.7), B220-PE (CD45RA-14.8), Annexin V-APC, CD45-V500, CD14-V450, IgG2a-PE rat isotype (R35–95), IgG2a-APC rat isotype), IgG2a-FITC, IgG2a-V500, IgG2a-V450 rat isotype. CD44-PE (IM7) was purchased from Biolegend, San Diego, CA, USA. Thymocytes, splenocytes and whole blood cells (1 × 106 cells) were incubated with fluorochrome-conjugated antibodies (0.5 μg) for 30 minutes on ice, avoiding light. After two PBS washes, cells were analysed by flow cytometry using BD FACSAria (BD Biosciences) flow cytometer and BD FACSDiva software (BD Biosciences), as previously described76 (link). Splenic B-cells (B220+), splenic T-cells (CD3+), splenic CD4+ and CD8+ cells were sorted by antigenic criteria using BD FACSAria flow cytometer/cell sorter, with a purity grade of 99%.
Fiume G., Scialdone A., Albano F., Rossi A., Maria Tuccillo F., Rea D., Palmieri C., Caiazzo E., Cicala C., Bellevicine C., Falcone C., Vecchio E., Pisano A., Ceglia S., Mimmi S., Iaccino E., Laurentiis A.D., Pontoriero M., Agosti V., Troncone G., Mignogna C., Palma G., Arra C., Mallardo M., Maria Buonaguro F., Scala G, & Quinto I. (2015). Impairment of T cell development and acute inflammatory response in HIV-1 Tat transgenic mice. Scientific Reports, 5, 13864.
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5

Multiparametric Phenotyping of Stromal Cells

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This section describes the experiment shown in Figure 4A. HS-5 and NKTert cells were harvested with PBS supplemented with 0.02% EDTA to avoid cleavage of surface antigens. Cells were stained with the following antibody conjugates: CD146-BV421 (clone P1H12; Biolegend), CD31-BV650 (M89D3; BD Biosciences), CD105-FITC (43A3; Biolegend), CD90-PerCP/Cy5.5 (5E10; BD Biosciences), CD44-PE (IM7; Biolegend), podoplanin-PE/Dazzle (NC-08; Biolegend), CD271-PE/Cy7 (ME20.4-1.H4; Miltenyi Biotec), CD26-APC (M-A261; BD Biosciences), CD73-APC/Cy7 (AD2; Biolegend) as well as Fixable Viability Dye eFlour 506 (ThermoFisher Scientific). Cells were analyzed on a LSR Fortessa flow cytometer (BD Biosciences). Results are shown as fluorescence intensity histograms gated on live singlets.
Herbst S.A., Stolarczyk M., Becirovic T., Czernilofsky F., Liu Y., Kolb C., Knoll M., Herling M., Müller-Tidow C, & Dietrich S. (2021). Phagocytosis by stroma confounds coculture studies. iScience, 24(9), 103062.
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6

Flow Cytometry Analysis of Bone Marrow and Spleen Cells

Cited 1 time
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Bone marrow and spleen cells were stained for flow cytometry as previously described (Mayer et al., 2017 (link)). Gating strategy for BM erythroid cells was described in the Supplementary Figure 1. Cells were stained with the following antibodies: CD45-FITC (30-F11, eBioscience), CD11b-FITC (M1/70, eBioscience), Gr-1-FITC (RB6-8C5, eBioscience), Ter119-APC (TER119, Biolegend) and CD44-PE (IM7, Biolegend), F4/80-APC (BM8, Biolegend), Gr-1-FITC (RB6-8C5, Biolegend), CD3-PE (145-2C11, Biolegend), Ly6C-PE (HK1.4, Biolegend), CD106-PE (429, Biolegend), CD45R/B220-APC (RA3.682, Biolegend). Cells were analyzed using AccuriC6 flow cytometer and data were analyzed using FlowJo software (v10.7.1).
Myneni V.D., Szalayova I, & Mezey E. (2021). Differences in Steady-State Erythropoiesis in Different Mouse Bones and Postnatal Spleen. Frontiers in Cell and Developmental Biology, 9, 646646.
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7

Multiparameter Flow Cytometry for Immune Cell Profiling

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Single-cell suspensions in FACs buffer were incubated in Fc block (2.4G2, BD Biosciences, Franklin Lakes, NJ, USA) for 15 min then samples were washed in FACs buffer and centrifuged at 400 × g for 4 min. Cells were then incubated in a solution containing fluorescently labelled antibodies for 30 min in the dark on ice. Following staining, flow cytometry was performed on a BD FACS Canto II (BD Biosciences) or a LSRFortessa™ (BD Biosciences). Analysis was performed on FlowJo software (BD Biosciences) using the gating strategy outlined in Additional file 2: Fig S2.
The following antibodies were used for the detection and phenotyping of immune cells: CD45-BV510 (30-F11; Biolegend, San Diego, CA, USA), CD3-APC (17A2; Biolegend), B220-APC-Cy7 (RA3-6B2; Biolegend), CD4-BV421 (RM4-5; Biolegend), CD8a-PerCP-Cy5.5 (53–6.7; Biolegend), CD62L-FITC (MEL-14; Biolegend), CD44-PE (IM7; Biolegend), CD11b-PE-Cy7 (M1/70; Biolegend), Ly6G-APC (1A8; Biolegend), Ly6C-PE (HK1.4; Biolegend), and CD49d-AF488 (R1-2; Biolegend).
Peck T., Davis C., Lenihan-Geels G., Griffiths M., Spijkers-Shaw S., Zubkova O.V, & La Flamme A.C. (2023). The novel HS-mimetic, Tet-29, regulates immune cell trafficking across barriers of the CNS during inflammation. Journal of Neuroinflammation, 20, 251.
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