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Ab55138

Manufactured by Abcam
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Sourced in United Kingdom

Ab55138 is a laboratory equipment product manufactured by Abcam. It is designed for general laboratory use. No further details about its core function or intended use are provided.

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Lab products found in correlation

Cell activation cocktail, by BioLegend (2 mentions) Foxp3 fjk 16s staining, by Thermo Fisher Scientific (2 mentions) Cytofix cytoperm solution, by BD (2 mentions) Ab93434, by Abcam (1 mentions) Ab113509, by Abcam (1 mentions) A21285, by Thermo Fisher Scientific (1 mentions) Superfrost slide, by Thermo Fisher Scientific (1 mentions) Axio imager m1 microscope, by Zeiss (1 mentions) Axiocam mrm camera, by Zeiss (1 mentions) Dm5500b, by Leica Microsystems (1 mentions)

4 protocols using ab55138

1

Immunofluorescence Analysis of Corneal Proteins

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Excised mouse eyes or human donor cornea button were fixed in 4% paraformaldehyde in 0.1 mol/L phosphate buffer (pH 7.4) at 4 °C overnight and paraffin embedded. Five-micrometer sections were then mounted on Super Frost slides (Fisher Scientific). The sections were de-paraffinized and hydrated in a graded ethanol series (100%, 95% and 70% and 50% ethanol and ddH2O for 5 min each) and subject to antigen retrieval in 10 mM Na-citrate, then blocked with 2% BSA in PBS and incubated overnight at 4 °C with 1st antibodies. After washes in PBS, slides were incubated with Fluor-conjugated 2nd antibody for one hour and washed. Sections were mounted with prolong anti-fade mounting reagent with DAPI (Molecular Probes, Life Technologies) and imaged with AxioImager M1 microscope with AxioCam MRm camera (Zeiss).
The following antibodies were used: Rabbit polyclonal anti-Glutaminase (GLS1) antibody 1:200 (ab93434, Abcam); Rabbit anti-GLS2 antibody 1:200 (ab113509, Abcam); Mouse monoclonal anti-GGT1 antibody 1:200 (ab55138, Abcam); Rabbit anti-ZO1 1:200 (402200, Life Technologies); Mouse anti-ZO1 1:200 (339100, Invitrogen); Rabbit anti-Nitrotyrosine 1:200 (A-21285, Thermo Scientific); Secondary Alexa-488 and Alexa-568 antibodies (Molecular Probes) were used at 1:200 concentrations. Mean intensity (MFI) quantification of endothelium was conducted using ImageJ.
Zhang W., Li H., Ogando D.G., Li S., Feng M., Price FW J.r., Tennessen J.M, & Bonanno J.A. (2017). Glutaminolysis is Essential for Energy Production and Ion Transport in Human Corneal Endothelium. EBioMedicine, 16, 292-301.
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2

Multiparametric Flow Cytometry Analysis

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Single-cell suspensions were incubated with Fc receptor-blocking anti- CD16/CD32 antibodies (2.4G2) for 15 min at 4 °C. Cell suspensions were next stained with surface fluorescent-conjugated antibodies for 30 min at 4 °C. The following antibodies were used: CD3 (17A2), CD4 (GK1.5), CD8 (53-6.7), CD62L (MEL-14), CD44 (1M7), CD25 (3C7), CD11b (M1/70), CD45 (30-F11), and MHC II (M5/114.15.2). For surface labeling of GGT1, unconjugated mouse monoclonal anti-GGT1 (1:100; ab55138, Abcam) was incubated with surface markers for 1 h at 4 °C. Then FITC-conjugated species-specific secondary antibody (1:100) was added for 1 h at 4 °C. For intracellular staining of IFN-γ and IL-17A, T cells were stimulated for 4 h with Cell Activation Cocktail (BioLegend). Cells were then fixed with Cytofix/Cytoperm solution (BD Bioscience) and stained with antibodies to IL-17A (TC11-18H10.1), or IFN-γ (XMG1.2). Foxp3 (FJK-16S) staining was performed according to the manufacturer’s protocol (eBioscience). Cells were analyzed by flow cytometry on an LSRII (BD Biosciences) with FlowJo software (Tree Star Inc.). Antibodies were purchased from BioLegend, BD Biosciences, or eBioscience and were used at 1:300 dilution unless stated otherwise.
Mendiola A.S., Ryu J.K., Bardehle S., Meyer-Franke A., Ang K.K., Wilson C., Baeten K.M., Hanspers K., Merlini M., Thomas S., Petersen M.A., Williams A., Thomas R., Rafalski V.A., Meza-Acevedo R., Tognatta R., Yan Z., Pfaff S.J., Machado M.R., Bedard C., Coronado P.E., Jiang X., Wang J., Pleiss M.A., Green A.J., Zamvil S.S., Pico A.R., Bruneau B.G., Arkin M.R, & Akassoglou K. (2020). Transcriptional profiling and therapeutic targeting of oxidative stress in neuroinflammation. Nature immunology, 21(5), 513-524.
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3

Oral Cancer Tissue Analysis

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Frozen tissues of 15 patients with oral cancer were analysed to assess sensitivity and specificity. Normal tissue of the oral cavity of each patient was used as a negative control. Tissues were obtained from the Erasmus Medical Center (Rotterdam, The Netherlands) tissue bank. Cryosections were cut into slices of 10 μm. The cryosections were imaged by fluorescence microscopy (Leica microsystems DM5500 B, Eindhoven, The Netherlands) at 0, 1, 5 and 10 min after application of gGlu-HMRG (50 μl of 50 μM in PBS). Fluorescence intensity (negative, weak, or positive) was scored independently by two authors (MS & HH). Subsequently, each slice was stained using an anti-GGT1 antibody (dilution 1:800; ab55138, Abcam, Cambridge, UK). The staining of epithelium was also scored (negative, weak, or positive) by two authors (LH & HH). Concordance between fluorescence and GGT status was subsequently evaluated.
Slooter M.D., Handgraaf H.J., Boonstra M.C., van der Velden L.A., Bhairosingh S.S., Que I., de Haan L.M., Keereweer S., van Driel P.B., Chan A., Kobayashi H., Vahrmeijer A.L, & Löwik C.W. (2018). Detecting tumour-positive resection margins after oral cancer surgery by spraying a fluorescent tracer activated by gamma-glutamyltranspeptidase. Oral oncology, 78, 1-7.
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4

Multiparametric Flow Cytometry Analysis

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Single-cell suspensions were incubated with Fc receptor-blocking anti- CD16/CD32 antibodies (2.4G2) for 15 min at 4 °C. Cell suspensions were next stained with surface fluorescent-conjugated antibodies for 30 min at 4 °C. The following antibodies were used: CD3 (17A2), CD4 (GK1.5), CD8 (53-6.7), CD62L (MEL-14), CD44 (1M7), CD25 (3C7), CD11b (M1/70), CD45 (30-F11), and MHC II (M5/114.15.2). For surface labeling of GGT1, unconjugated mouse monoclonal anti-GGT1 (1:100; ab55138, Abcam) was incubated with surface markers for 1 h at 4 °C. Then FITC-conjugated species-specific secondary antibody (1:100) was added for 1 h at 4 °C. For intracellular staining of IFN-γ and IL-17A, T cells were stimulated for 4 h with Cell Activation Cocktail (BioLegend). Cells were then fixed with Cytofix/Cytoperm solution (BD Bioscience) and stained with antibodies to IL-17A (TC11-18H10.1), or IFN-γ (XMG1.2). Foxp3 (FJK-16S) staining was performed according to the manufacturer’s protocol (eBioscience). Cells were analyzed by flow cytometry on an LSRII (BD Biosciences) with FlowJo software (Tree Star Inc.). Antibodies were purchased from BioLegend, BD Biosciences, or eBioscience and were used at 1:300 dilution unless stated otherwise.
Mendiola A.S., Ryu J.K., Bardehle S., Meyer-Franke A., Ang K.K., Wilson C., Baeten K.M., Hanspers K., Merlini M., Thomas S., Petersen M.A., Williams A., Thomas R., Rafalski V.A., Meza-Acevedo R., Tognatta R., Yan Z., Pfaff S.J., Machado M.R., Bedard C., Coronado P.E., Jiang X., Wang J., Pleiss M.A., Green A.J., Zamvil S.S., Pico A.R., Bruneau B.G., Arkin M.R, & Akassoglou K. (2020). Transcriptional profiling and therapeutic targeting of oxidative stress in neuroinflammation. Nature immunology, 21(5), 513-524.
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