mouse anti human cxcr6 monoclonal antibody  (R&D Systems)

Journal: Scientific Reports

Article Title: Higher CXCL16 exodomain is associated with aggressive ovarian cancer and promotes the disease by CXCR6 activation and MMP modulation

doi: 10.1038/s41598-019-38766-6

Figure Lengend Snippet: CXCR6 expression in ovarian cancer tissues and cell lines. Ovarian cancer tissues from normal (n = 12), endometrioid (n = 27) and serous (n = 33) were probed with antibody against CXCR6. CXCR6 was developed using DAB (brown). Images (40X) were captured using TissueFAXS cell analysis system from tissuegnostics (Panel A). Immuno-intensity of CXCR6 as shown in box plot was quantified using Histoquest image analysis software (Panel B). Line in the middle represents median value. ★ p < 0.05 when compared to control; ✜ p < 0.05 when compared between the endometrioid and serous papillary groups. CXCR6 protein expression in OvCa and normal epithelial cells was confirmed by FACS analysis. Cells were stained for CXCR6 expression using a purified phycoerythrin-conjugated anti-human CXCR6 (Panel C).

Article Snippet: After blocking, TMA was washed with Tris-buffer, and incubated for 1 hour in humidity chamber at RT with mouse anti-human-CXCR6 monoclonal antibody (1:75 dilution of 1.0 μg/ml stock) (R&D Systems).

Techniques: Expressing, Cell Analysis, Software, Control, Staining, Purification

Journal: Scientific Reports

Article Title: Higher CXCL16 exodomain is associated with aggressive ovarian cancer and promotes the disease by CXCR6 activation and MMP modulation

doi: 10.1038/s41598-019-38766-6

Figure Lengend Snippet: CXCR6-CXCL16 axis promotes ovarian cancer cell migration and invasion. Ovarian cancer cells (OVCAR-3 and SKOV-3) were allowed to migrate (Panel –A,B) or invade (Panel –C,D) in tumor migration and invasion chamber under the chemotactic gradient of CXCL16 (100 ng/ml) with or without anti-CXCR6 antibody (1.0 μg/ml) blocking of CXCR6. Cells that had migrated or invaded through the membrane or matrigel were fixed, stained and counted. Bars represent the mean ± standard error (n = 20) of number of migrated or invaded cells in response to CXCL16. ★★ p < 0.01 represent statistically significant difference between cells migrating/invading towards CXCL16 gradient compared to control and ✜✜ p < 0.01 represent statistically significant difference between cells migrating/invading towards CXCL16 gradient with and without CXCR6 blockade.

Article Snippet: After blocking, TMA was washed with Tris-buffer, and incubated for 1 hour in humidity chamber at RT with mouse anti-human-CXCR6 monoclonal antibody (1:75 dilution of 1.0 μg/ml stock) (R&D Systems).

Techniques: Migration, Blocking Assay, Membrane, Staining, Control

Journal: PLoS ONE

Article Title: CXCL16 and CXCR6 Are Coexpressed in Human Lung Cancer In Vivo and Mediate the Invasion of Lung Cancer Cell Lines In Vitro

doi: 10.1371/journal.pone.0099056

Figure Lengend Snippet: Immunohistochemistry was performed to determine the protein expression of CXCL16-CXCR6 and CXCL12-CXCR4 in human primary lung cancer tissues. The antibodies used in this experiments were mouse anti-human CXCR4 (25 µg/ml), mouse anti-human CXCR6 (25 µg/ml), mouse anti-human CXCL12 (20 µg/ml) and goat anti-human CXCL16 (20 µg/ml) antibodies. It was demonstrated in Fig. 1 that a specific brown-coloured staining for CXCL16-CXCR6 and CXCL12-CXCR4 in the cytoplasm and membrane of human different pathological types of lung cancer cells. Moderate-to-strong, brown-coloured staining for CXCL16 and CXCR6 was also observed in normal lung tissues, but the positive expression was mainly restricted to the alveolar epithelial cells and inflammatory cells. There was no evidence for nonspecific staining with the control antibody. The pictures were the representative of the experiments. AC: adenocarcinomas; SC: squamous carcinomas; BC: bronchoalveolar carcinoma; Con: normal lung tissues; Villi: human first-trimester villous tissues, as a positive control. Magnification, ×200.

Article Snippet: After blocking with 10% FBS, the recovered cells were incubated with mouse anti-human PE (phycoerythrin) -CXCR6 monoclonal antibody (1∶10; R&D Systems, Inc), mouse anti-human PE-CY5-CXCR4 monoclonal antibody (1∶5; eBioscience), mouse PE-IgG2B isotype (R&D Systems, Inc) or mouse PE-CY5-IgG2a isotype (eBioscience), in the recommended usage for 1 h at room temperature in darkness.

Techniques: Immunohistochemistry, Expressing, Staining, Membrane, Control, Positive Control

Journal: PLoS ONE

Article Title: CXCL16 and CXCR6 Are Coexpressed in Human Lung Cancer In Vivo and Mediate the Invasion of Lung Cancer Cell Lines In Vitro

doi: 10.1371/journal.pone.0099056

Figure Lengend Snippet: Expression intensity of CXCL16/CXCR6 and CXCL12/CXCR4 protein in human lung cancer tissues.

Article Snippet: After blocking with 10% FBS, the recovered cells were incubated with mouse anti-human PE (phycoerythrin) -CXCR6 monoclonal antibody (1∶10; R&D Systems, Inc), mouse anti-human PE-CY5-CXCR4 monoclonal antibody (1∶5; eBioscience), mouse PE-IgG2B isotype (R&D Systems, Inc) or mouse PE-CY5-IgG2a isotype (eBioscience), in the recommended usage for 1 h at room temperature in darkness.

Techniques: Expressing

Journal: PLoS ONE

Article Title: CXCL16 and CXCR6 Are Coexpressed in Human Lung Cancer In Vivo and Mediate the Invasion of Lung Cancer Cell Lines In Vitro

doi: 10.1371/journal.pone.0099056

Figure Lengend Snippet: Immunocytochemistry was conducted to detect the protein expression of CXCL16-CXCR6 and CXCL12-CXCR4 in human lung cancer cell lines. The antibodies used in this experiments were mouse anti-human CXCR4 (25 µg/ml), mouse anti-human CXCR6 (25 µg/ml), mouse anti-human CXCL12 (20 µg/ml) and goat anti-human CXCL16 (20 µg/ml) antibodies. Positive brown-coloured staining for both CXCL16 and CXCR6 was clearly observed in the cytoplasm and cytomembrane of A549, H292 and 95D cells, respectively. Furthermore, CXCL12 and CXCR4 were also co-expressed in the three lung cancer cell lines. No background staining was observed in the isotype control. A: The images were representative of the experiments; B: The relative expression intensity of CXCL16-CXCR6 and CXCL12-CXCR4 in A549, H292 and 95D cells. Tro: human primary cultured trophoblast cells as a positive control. Magnification, ×200.

Article Snippet: After blocking with 10% FBS, the recovered cells were incubated with mouse anti-human PE (phycoerythrin) -CXCR6 monoclonal antibody (1∶10; R&D Systems, Inc), mouse anti-human PE-CY5-CXCR4 monoclonal antibody (1∶5; eBioscience), mouse PE-IgG2B isotype (R&D Systems, Inc) or mouse PE-CY5-IgG2a isotype (eBioscience), in the recommended usage for 1 h at room temperature in darkness.

Techniques: Immunocytochemistry, Expressing, Staining, Control, Cell Culture, Positive Control

Journal: PLoS ONE

Article Title: CXCL16 and CXCR6 Are Coexpressed in Human Lung Cancer In Vivo and Mediate the Invasion of Lung Cancer Cell Lines In Vitro

doi: 10.1371/journal.pone.0099056

Figure Lengend Snippet: Flow cytometry (FCM) was used to detect the membrane expression of CXCR6 in lung cancer cell lines. The cells of 1×10 5 were counted and the dilution ratio of (phycoerythrin)-CXCR6 monoclonal antibody and PE-CY5-CXCR4 monoclonal antibody were 1∶10 and 1∶5, respectively. The histogram demonstrated the average expression proportion of CXCR6 and CXCR4 in A549, H292 and 95D cells, respectively. The FCM pictures were representative of the experiments. The percentage of membrane CXCR6-positive cells in A549, H292 and 95D was 52.4±5.80, 56.03±11.42 and 34.8±6.17, respectively. Furthermore, the membrane expression of CXCR4 was also observed in A549, H292 and 95D cells at the same time. The experiments were repeated three times and the images were representative of the experiments. Error bars depict the standard error of the mean.

Article Snippet: After blocking with 10% FBS, the recovered cells were incubated with mouse anti-human PE (phycoerythrin) -CXCR6 monoclonal antibody (1∶10; R&D Systems, Inc), mouse anti-human PE-CY5-CXCR4 monoclonal antibody (1∶5; eBioscience), mouse PE-IgG2B isotype (R&D Systems, Inc) or mouse PE-CY5-IgG2a isotype (eBioscience), in the recommended usage for 1 h at room temperature in darkness.

Techniques: Flow Cytometry, Membrane, Expressing

Journal: PLoS ONE

Article Title: CXCL16 and CXCR6 Are Coexpressed in Human Lung Cancer In Vivo and Mediate the Invasion of Lung Cancer Cell Lines In Vitro

doi: 10.1371/journal.pone.0099056

Figure Lengend Snippet: To silence the CXCR6 gene expression, we transfected A549 cells with phU6/GFP/Neo plasmid containing short hairpin RNA (shRNA) molecules targeted against CXCR6, with the usage of Lipofectamine 2000 (Invitrogen). The sequences for three shRNA oligonucleotides were: (CXCR6-2819-1): 5′-ctGAG GAC AAT TCC AAG ACT T-3′ (sense) and 5′-AAG TCT TGG AAT TGT CCT CAG-3′ (Anti-sense); (CXCR6-2820-2) 5′-ctCAC CAT GAT TGT CTG CTA T-3′ (sense) and 5′-ATA GCA GAC AAT CAT GGT GAG-3′ (Anti-sense); (CXCR6-2821-1) 5′-gcTTG CTC ATC TGG GTG ATA T-3′ (sense) and 5′-ATA TCA CCC AGA TGA GCA AGC-3′ (Anti-sense). Efficiency of RNA interference against CXCR6 was validated by western blot. It was shown in Fig. 7 that CXCR6 protein was substantially expressed in A549 cells (bank-control, without any treatment) and the RNA interference technology effectively inhibited CXCR6 expression in A549 cells. Lane 1: Blank-control; Lane 2: RNA-control; Lane 3: CXCR6 shRNA-(2821-1); Lane 4: CXCR6 shRNA-(2820-2); Lane 5: CXCR6 shRNA-(2819-1).

Article Snippet: After blocking with 10% FBS, the recovered cells were incubated with mouse anti-human PE (phycoerythrin) -CXCR6 monoclonal antibody (1∶10; R&D Systems, Inc), mouse anti-human PE-CY5-CXCR4 monoclonal antibody (1∶5; eBioscience), mouse PE-IgG2B isotype (R&D Systems, Inc) or mouse PE-CY5-IgG2a isotype (eBioscience), in the recommended usage for 1 h at room temperature in darkness.

Techniques: Gene Expression, Transfection, Plasmid Preparation, shRNA, Western Blot, Control, Expressing

Journal: PLoS ONE

Article Title: CXCL16 and CXCR6 Are Coexpressed in Human Lung Cancer In Vivo and Mediate the Invasion of Lung Cancer Cell Lines In Vitro

doi: 10.1371/journal.pone.0099056

Figure Lengend Snippet: The MTT assay was applied to evaluate the effects of CXCL16-CXCR6 on cell viability in vitro . A: A549 cells from blank-control, shRNA-control and CXCR6-shRNA (2819-1) groups were seeded in 96-well flat-bottom microplates with a density of 2×10 3 cells/well. The in vitro viability was observed at 24, 48, 72, 96 and 120 h respectively. B: After starved with 1640 without FCS for 12 h, the cells from blank-control, shRNA-control or CXCR6-shRNA (2819-1) groups were treated with control medium (1640) or CXCL16 at a concentration of 100 ng/mL for 48 h. Then the in vitro viability was recorded by MTT assay. The experiments were repeated three times. Error bars depict the standard error of the mean. a P<0.01, compared with the corresponding control.

Article Snippet: After blocking with 10% FBS, the recovered cells were incubated with mouse anti-human PE (phycoerythrin) -CXCR6 monoclonal antibody (1∶10; R&D Systems, Inc), mouse anti-human PE-CY5-CXCR4 monoclonal antibody (1∶5; eBioscience), mouse PE-IgG2B isotype (R&D Systems, Inc) or mouse PE-CY5-IgG2a isotype (eBioscience), in the recommended usage for 1 h at room temperature in darkness.

Techniques: MTT Assay, In Vitro, Control, shRNA, Concentration Assay

Journal: PLoS ONE

Article Title: CXCL16 and CXCR6 Are Coexpressed in Human Lung Cancer In Vivo and Mediate the Invasion of Lung Cancer Cell Lines In Vitro

doi: 10.1371/journal.pone.0099056

Figure Lengend Snippet: Invasion assay was employed to investigate the effects of CXCL16-CXCR6 axis on the invasive ability of A549, 95D and H292 cells in vitro . Firstly, the isolated A549, H292 or 95D cells (1×10 5 /200 µl serum-free 1640) were plated in the upper chamber, and treated with CM, CXCL16 (100 ng/ml) or a combination of CM or CXCL16 with CXCL16 neutralization antibody(100 ng/mL). Secondly, the A549 cells, from the blank-control, phU6/GFP/Neo-CXCR6 and phU6/GFP/Neo group, were seeded on the upper chamber at a density of (1×10 5 /200 µl serum-free 1640), then treated with CXCL16 (100 ng/ml) or CM. The cells migrated to the lower surface were counted and the invasive index was calculated as the proportion of the migrated cells of the experiment group to that of its own control. Error bars depict the standard error of the mean. Con: the control; CXCL16: treated with 100 ng/ml CXCL16; antiCXCL16: treated with 100 ng/mL CXCL16 neutralizing antibody; CM: conditioned medium for A549, 95D or H292; Blank-control: without any treatment, shRNA-control: phU6/GFP/Neo; CXCR6-shRNA: phU6/GFP/Neo-CXCR6 (2819-1). a P<0.01 compared to the vehicle control; b P<0.05, c P<0.01, compared to the CXCL6 alone; d P<0.05, e P<0.01compared to the CM treatment group.

Article Snippet: After blocking with 10% FBS, the recovered cells were incubated with mouse anti-human PE (phycoerythrin) -CXCR6 monoclonal antibody (1∶10; R&D Systems, Inc), mouse anti-human PE-CY5-CXCR4 monoclonal antibody (1∶5; eBioscience), mouse PE-IgG2B isotype (R&D Systems, Inc) or mouse PE-CY5-IgG2a isotype (eBioscience), in the recommended usage for 1 h at room temperature in darkness.

Techniques: Invasion Assay, In Vitro, Isolation, Neutralization, Control, shRNA

Journal: PLoS ONE

Article Title: Distinct Roles for CXCR6 + and CXCR6 − CD4 + T Cells in the Pathogenesis of Chronic Colitis

doi: 10.1371/journal.pone.0065488

Figure Lengend Snippet: ( A, B ) mRNA expression of CXCL16 ( A ) and CXCR6 ( B ) was evaluated by Q-PCR. The expression of CXCL16 and CXCR6 is higher in colonic mucosa of patients with Crohn’s disease (CD) compared with patients with ulcerative colitis (UC) and healthy controls. Data were normalized to expression of GAPDH mRNA. ( n = 5–10; mean and s.e.m.). *, P < 0.05. ( C ) Correlation between CXCL16 and CXCR6 mRNA expression in the colonic mucosa of CD patients. Statistical analysis was performed by Spearman’s correlation; correlation coefficient = 0.76, P = 0.024. ( D, E ) Immunohistochemistry of CXCL16 ( D ) and CXCL16 ( E ) was performed on colonic mucosa of patients with CD and healthy controls. CXCL16 positive staining was observed on epithelial cells and a subset of colonic LP cells in CD patients ( D ). CXCR6 was strongly expressed by small round cells in CD mucosa ( E ). Scale bars, 50 µm, IgG indicates a control antibody. Representative photomicrographs obtained from the analysis of five or six specimens per group are shown.

Article Snippet: The sections were incubated with 5% bovine serum albumin in PBS for 30 min at room temperature and then with goat anti-human CXCL16 polyclonal Ab (R&D Systems), mouse anti-human CXCR6 monoclonal Ab (R&D Systems), or an identical concentration of control goat or mouse IgG, overnight at 4°C.

Techniques: Expressing, Immunohistochemistry, Staining, Control

Journal: PLoS ONE

Article Title: Distinct Roles for CXCR6 + and CXCR6 − CD4 + T Cells in the Pathogenesis of Chronic Colitis

doi: 10.1371/journal.pone.0065488

Figure Lengend Snippet: ( A ) Cxcl16 mRNA levels in colonic epithelium (CEC) and distal colon tissues were analyzed by Q-PCR 8 weeks after transfer. The expression of Cxcl16 mRNA increased in both epithelium and colon tissues of the transfer model compared with healthy Rag1 −/− mice. Data were normalized to expression of Gapdh mRNA. ( n = 5; mean and s.d.). **, P < 0.01. ( B ) CXCL16 immunostaining of the distal colon in colitic and healthy Rag1 −/− mice. Scale bars, 100 µm. Data are representative of two independent experiments. ( C ) CXCR6 expression on CD4 + T cells was analyzed by flow cytometry using a mouse CXCL16-human IgG-Fc fusion protein or control human IgG-Fcγ at 8-week post transfer. CXCR6 was expressed at high levels by the majority of colonic LP CD4 + T cells in the colitic mice, by about half of the SP and MLN CD4 + T cells, and by ∼20% of BM cells. ( D ) Absolute numbers of CXCR6 + and CXCR6 − CD4 + T cells in each of tissues were calculated based on the flow cytometric analysis described in ( C ). Data are representative of three independent experiments (mean and s.d.). **, P < 0.01.

Article Snippet: The sections were incubated with 5% bovine serum albumin in PBS for 30 min at room temperature and then with goat anti-human CXCL16 polyclonal Ab (R&D Systems), mouse anti-human CXCR6 monoclonal Ab (R&D Systems), or an identical concentration of control goat or mouse IgG, overnight at 4°C.

Techniques: Expressing, Immunostaining, Flow Cytometry, Control

Journal: PLoS ONE

Article Title: Distinct Roles for CXCR6 + and CXCR6 − CD4 + T Cells in the Pathogenesis of Chronic Colitis

doi: 10.1371/journal.pone.0065488

Figure Lengend Snippet: LP CXCR6 − and CXCR6 + cells were analyzed for expression of activation and memory markers at week 8 post-transfer of naïve CD4 + T cells. ( A – C ) CD4 + T cells were gated as CD127 − CD62L − CD27 − CD43 + CD44 + to measure the proportion of effector T cells (a). ( D – F ) The effector memory population (CD44 + CD127 + ) was subdivided using CD62L and CD27 to measure early effector memory cells (CD62L − CD27 + CD43 + , b) and late effector memory cells (CD62L − CD27 − CD43 + , c). Data are representative of three independent experiments. ( G ) The relative percentages of effector, early effector memory and late effector memory in each subset are shown in a pie chart.

Article Snippet: The sections were incubated with 5% bovine serum albumin in PBS for 30 min at room temperature and then with goat anti-human CXCL16 polyclonal Ab (R&D Systems), mouse anti-human CXCR6 monoclonal Ab (R&D Systems), or an identical concentration of control goat or mouse IgG, overnight at 4°C.

Techniques: Expressing, Activation Assay

Journal: PLoS ONE

Article Title: Distinct Roles for CXCR6 + and CXCR6 − CD4 + T Cells in the Pathogenesis of Chronic Colitis

doi: 10.1371/journal.pone.0065488

Figure Lengend Snippet: Intracellular staining for cytokine and transcription factors in CD4 + T cells was performed 8 weeks after naïve CD4 + T-cell transfer. ( A, B ) The frequency of IL-2 + cells and IFN-γ + cells was analyzed in the CXCR6 − (upper) and CXCR6 + (lower) subsets ( A ) and the absolute number of T cells were graphed on the basis of the flow cytometric analysis ( B ). ( C, D ) The frequency of IL-17A + cells and TNF-α + cells was analyzed in CXCR6 − (upper) and CXCR6 + (lower) subset ( C ) and the numbers were graphed on the basis of flow cytometric analysis ( D ). ( E ) The frequency of IFN-γ + cells and IL-17A + cells was analyzed. ( F ) The frequency of T-bet + cells and RORγt + cells was analyzed. All data are representative from four independent experiments (mean and s.d.). *, P < 0.05 **, P < 0.01 ***, P < 0.001.

Article Snippet: The sections were incubated with 5% bovine serum albumin in PBS for 30 min at room temperature and then with goat anti-human CXCL16 polyclonal Ab (R&D Systems), mouse anti-human CXCR6 monoclonal Ab (R&D Systems), or an identical concentration of control goat or mouse IgG, overnight at 4°C.

Techniques: Staining

Journal: PLoS ONE

Article Title: Distinct Roles for CXCR6 + and CXCR6 − CD4 + T Cells in the Pathogenesis of Chronic Colitis

doi: 10.1371/journal.pone.0065488

Figure Lengend Snippet: ( A ) Body weight of Rag1 −/− recipients of i.v. injected purified CD45RB high CD4 + T cells form Cxcr6 +/Egfp or Cxcr6 Egfp/Egfp (CXCR6-deficient) mice on day 0, presented as percent of original weight. ( B ) Colon weight of the mice in ( A ) on week 7. Data are representative of two independent experiments (mean and s.d.). ( C ) Histology of colon tissues from the mice in B . ( D ) CXCR6 expression by LP CD4 + T cells was analyzed by flow cytometry using CXCL16-hFc at 7-week post transfer. ( E ) Expression levels of indicated cytokines in distal colon were analyzed by Q-PCR at 7 weeks after the transfer. Data were normalized to expression of Gapdh . ( n = 4 or 5; mean and s.d.).

Article Snippet: The sections were incubated with 5% bovine serum albumin in PBS for 30 min at room temperature and then with goat anti-human CXCL16 polyclonal Ab (R&D Systems), mouse anti-human CXCR6 monoclonal Ab (R&D Systems), or an identical concentration of control goat or mouse IgG, overnight at 4°C.

Techniques: Injection, Purification, Expressing, Flow Cytometry

Journal: PLoS ONE

Article Title: Distinct Roles for CXCR6 + and CXCR6 − CD4 + T Cells in the Pathogenesis of Chronic Colitis

doi: 10.1371/journal.pone.0065488

Figure Lengend Snippet: ( A ) Schematic transfer protocol. An equal number of CD25 − CXCR6 − cells or CD25 − CXCR6 + CD4 + T cell isolated from colon LP of colitic mice was retransferred into Rag1 −/− mice. ( B ) Time course of changes in body weight after retransfer of the two subsets or transfer of CD45RB high naïve T cells. CXCR6 – transferred Rag1 −/− mice manifested progressive body weight loss to a similar extent to CD45RB high -transferred Rag1 −/− mice, whereas the cohort that received CXCR6 + cells did not show wasting. Data are expressed as the mean and s.e.m of two independent experiments. *, P < 0.05 ***, P < 0.001 versus CXCR6 + -transferred animals. ( C ) Histopathological analysis of distal colon tissue at 8 weeks post transfer. The transfer of CXCR6 − CD4 + T cells alone induced intestinal inflammation. Scale bars, 50 µm. ( D ) Number of CD4 + T cells in each group were calculated based on the flow cytometric analysis. ( E ) CXCR6 expression on CD4 + T cells in each group was analyzed by flow cytometry using CXCL16-hFc at 8-week post transfer. CXCR6 − CD4 + T cells express CXCR6 upon retransfer. ( F ) Intracellular cytokine staining was performed in LP CD4 + T cells in recipients of CXCR6 − CD4 + T cells at 8 weeks after transfer. The CXCR6 − subset produced IFN-γ and IL-17A coincident with the expression of CXCR6 upon retransfer. ( G ) LP CXCR6 − and CXCR6 + subsets were purified from CD45RB high -transferred Rag1 −/− mice at 8-week post transfer, labeled with CFSE and cultured with LP MHC class II + CD11c + cells at a 5∶1 ratio in the presence of 5 µg/ml anti-CD3ε Abs and 20 ng/ml IL-23 for 3 days. Proliferation was measured by CFSE dilution. Proliferation by CXCR6 − and CXCR6 + cells is depicted in the right columns. All data are representative from four independent experiments (mean and s.d.).

Article Snippet: The sections were incubated with 5% bovine serum albumin in PBS for 30 min at room temperature and then with goat anti-human CXCL16 polyclonal Ab (R&D Systems), mouse anti-human CXCR6 monoclonal Ab (R&D Systems), or an identical concentration of control goat or mouse IgG, overnight at 4°C.

Techniques: Isolation, Expressing, Flow Cytometry, Staining, Produced, Purification, Labeling, Cell Culture