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R&D Systems ng2 recombinant protein
Ng2 Recombinant Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ng2 recombinant protein/product/R&D Systems
Average 90 stars, based on 3 article reviews

ng2 recombinant protein - by Bioz Stars, 2026-03

90/100 stars

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(A) Expression of CSPG4 and gp100 on human melanoma cell lines was detected by flow cytometry using specific monoclonal antibodies and FITC-conjugated secondary antibody. Expression of CD146 was detected using FITC-conjugated mouse anti-human CD146. Gray lines: secondary antibody alone or isotype control; Black lines: anti- CSPG4, gp100 or CD146, as indicated. (B) 1G2 (HLA-A2+/ gp100154–162- reactive) T cell line was co-cultured with HLA-A2+ (M579-A2 and 624mel) or HLA-A2− (M171 and M579) melanoma cells. CSPG4 was detected on CD8+ lymphocytes by staining with specific monoclonal antibodies, followed by flow cytometry. (C) 2E2, 2C7 and 1H8 (all A2/MART-126–35- reactive) T cell clones were co-cultured and analyzed as described in B. (D) 1H8 T cell clone was co-cultured with melanoma cell lines as described in B. Presence of gp100 and CD146 on CD8+ cells was monitored by flow cytometry. One representative experiment of five performed is shown. (E) The four following cell lines were co-cultured: MART-126–35-reactive T cells (2D6), a non-relevant MUC-163–71-specific T cell line (line 33), melanoma cells (526mel) and MUC-163–71 peptide-pulsed T2 cells. CSPG4 and CD107A were detected on CD8+ lymphocytes by staining with specific monoclonal antibodies, followed by flow cytometry. To enable separate gating, the two lymphocyte populations 2D6 and line33 were pre-labeled with PKH67 and PKH26, respectively.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Lymphocyte imprinting with melanoma antigens acquired by trogocytosis facilitates identification of tumor-reactive T cells

doi: 10.4049/jimmunol.1202879

Figure Lengend Snippet: (A) Expression of CSPG4 and gp100 on human melanoma cell lines was detected by flow cytometry using specific monoclonal antibodies and FITC-conjugated secondary antibody. Expression of CD146 was detected using FITC-conjugated mouse anti-human CD146. Gray lines: secondary antibody alone or isotype control; Black lines: anti- CSPG4, gp100 or CD146, as indicated. (B) 1G2 (HLA-A2+/ gp100154–162- reactive) T cell line was co-cultured with HLA-A2+ (M579-A2 and 624mel) or HLA-A2− (M171 and M579) melanoma cells. CSPG4 was detected on CD8+ lymphocytes by staining with specific monoclonal antibodies, followed by flow cytometry. (C) 2E2, 2C7 and 1H8 (all A2/MART-126–35- reactive) T cell clones were co-cultured and analyzed as described in B. (D) 1H8 T cell clone was co-cultured with melanoma cell lines as described in B. Presence of gp100 and CD146 on CD8+ cells was monitored by flow cytometry. One representative experiment of five performed is shown. (E) The four following cell lines were co-cultured: MART-126–35-reactive T cells (2D6), a non-relevant MUC-163–71-specific T cell line (line 33), melanoma cells (526mel) and MUC-163–71 peptide-pulsed T2 cells. CSPG4 and CD107A were detected on CD8+ lymphocytes by staining with specific monoclonal antibodies, followed by flow cytometry. To enable separate gating, the two lymphocyte populations 2D6 and line33 were pre-labeled with PKH67 and PKH26, respectively.

Article Snippet: APC-conjugated-mouse anti human NG2/MCSP (CSPG4) (R&D Systems, Minneapolis, MN), mouse anti-human HMB45 (Serotec, Oxford, UK), FITC-conjugated mouse anti-human CD146, FITC-conjugated rat anti-human CD8 antibody (Serotec), FITC- or APC-conjugated mouse anti human CD8, PE or FITC conjugated mouse anti-human CD4, PE-conjugated mouse anti human IFN-γ, FITC- conjugated mouse anti-human CD107A (LAMP-1) (all from eBioscience, San Diego, CA), pacific blue-conjugated mouse anti-human CD107 (Biolegend), APC-mouse anti-human CD137 (BD Pharmingen), APC or DyLight 649-conjugated donkey anti-mouse IgG (Jackson, West Grove, PA) were purchased from the indicated companies.

Techniques: Expressing, Flow Cytometry, Bioprocessing, Control, Cell Culture, Staining, Clone Assay, Labeling

(A–C) Confocal microscopy of melanoma antigen-imprinted CD8+ T cell clones after co-incubation with HLA-A2-matched melanoma. (A) Capture of gp100; (B,C) Capture of CSPG4. (D) Co-culture of T cell clones with HLA-A2 mismatched melanoma.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Lymphocyte imprinting with melanoma antigens acquired by trogocytosis facilitates identification of tumor-reactive T cells

doi: 10.4049/jimmunol.1202879

Figure Lengend Snippet: (A–C) Confocal microscopy of melanoma antigen-imprinted CD8+ T cell clones after co-incubation with HLA-A2-matched melanoma. (A) Capture of gp100; (B,C) Capture of CSPG4. (D) Co-culture of T cell clones with HLA-A2 mismatched melanoma.

Techniques: Confocal Microscopy, Clone Assay, Incubation, Co-Culture Assay

PBMCs from melanoma patient 171 were cultured for 12 days in the presence of irradiated autologous melanoma (M171). Following, the cells were co-cultured with autologous (M171) or irrelevant (624mel) melanoma cells. CSPG4 was monitored on CD8+ lymphocytes by staining with specific monoclonal antibodies, followed by flow cytometry. (B) PBMCs from melanoma patient 579 were cultured for 12 days in the presence of irradiated autologous melanoma (M579). Following, the cells were co-cultured with autologous (M579) or irrelevant (M171) melanoma cells. IFN-γ secretion of non-stimulated and 12 day-stimulated cells after re-stimulation with irrelevant melanoma (624mel, empty bar) or autologous melanoma (M579, filled bar) was evaluated by ELISA. (C) Presence of CSPG4 was monitored on CD4+ lymphocytes derived from PBMCs (patient 579), 12 days after stimulation with autologous melanoma, by staining with specific monoclonal antibodies followed by flow cytometry.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Lymphocyte imprinting with melanoma antigens acquired by trogocytosis facilitates identification of tumor-reactive T cells

doi: 10.4049/jimmunol.1202879

Figure Lengend Snippet: PBMCs from melanoma patient 171 were cultured for 12 days in the presence of irradiated autologous melanoma (M171). Following, the cells were co-cultured with autologous (M171) or irrelevant (624mel) melanoma cells. CSPG4 was monitored on CD8+ lymphocytes by staining with specific monoclonal antibodies, followed by flow cytometry. (B) PBMCs from melanoma patient 579 were cultured for 12 days in the presence of irradiated autologous melanoma (M579). Following, the cells were co-cultured with autologous (M579) or irrelevant (M171) melanoma cells. IFN-γ secretion of non-stimulated and 12 day-stimulated cells after re-stimulation with irrelevant melanoma (624mel, empty bar) or autologous melanoma (M579, filled bar) was evaluated by ELISA. (C) Presence of CSPG4 was monitored on CD4+ lymphocytes derived from PBMCs (patient 579), 12 days after stimulation with autologous melanoma, by staining with specific monoclonal antibodies followed by flow cytometry.

Techniques: Cell Culture, Irradiation, Staining, Bioprocessing, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Derivative Assay

(A) TIL from melanoma patient 412 were cultured for 12 days in the presence of irradiated autologous melanoma. Expanded cells were co-cultured with HLA-A2-matched melanoma (624mel, M579-A2) or HLA-A2-mismatched melanoma cells (M171, M579). CSPG4 was monitored on CD8+ lymphocytes by staining with specific monoclonal antibodies followed by flow cytometry. One representative experiment of three performed is shown. (B) Melanoma antigens gp100 (in patients 1 and 2) and CSPG4 (in patient 3) were monitored on CD4+ and CD8+ TIL 4 days after tumor resection from the patient (the minimal time required for TIL expansion in the culture medium). Cells were stained with specific monoclonal antibodies followed by flow cytometry. Dot plots are the results of tumor antigen and background staining for each individual patient. Bars summarize the data from all three patients. Gray bars represent background staining with secondary antibody only (patient 1 and 2), or with isotype (patient 3). Black bars represent specific staining with anti-melanoma antigen specific Ab.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Lymphocyte imprinting with melanoma antigens acquired by trogocytosis facilitates identification of tumor-reactive T cells

doi: 10.4049/jimmunol.1202879

Figure Lengend Snippet: (A) TIL from melanoma patient 412 were cultured for 12 days in the presence of irradiated autologous melanoma. Expanded cells were co-cultured with HLA-A2-matched melanoma (624mel, M579-A2) or HLA-A2-mismatched melanoma cells (M171, M579). CSPG4 was monitored on CD8+ lymphocytes by staining with specific monoclonal antibodies followed by flow cytometry. One representative experiment of three performed is shown. (B) Melanoma antigens gp100 (in patients 1 and 2) and CSPG4 (in patient 3) were monitored on CD4+ and CD8+ TIL 4 days after tumor resection from the patient (the minimal time required for TIL expansion in the culture medium). Cells were stained with specific monoclonal antibodies followed by flow cytometry. Dot plots are the results of tumor antigen and background staining for each individual patient. Bars summarize the data from all three patients. Gray bars represent background staining with secondary antibody only (patient 1 and 2), or with isotype (patient 3). Black bars represent specific staining with anti-melanoma antigen specific Ab.

Techniques: Cell Culture, Irradiation, Staining, Bioprocessing, Flow Cytometry

MART-126–35-specific T cells (clone 2D6) were mixed with melanoma non-relevant, MUC-163–71-specific T cells (line 33) at the indicated ratios, and co-cultured with HLA A2-matched melanoma cells (526mel) or irrelevant melanoma cells (888mel). (A) The percentage of CSPG4+/CD8+ melanoma antigen (MA)-imprinted cells out of total CD8+ cells was correlated to the proportion of melanoma antigen-specific cells in the culture (MA-specific cells, R2=0.99, p<0.0001) (left panel). A representative dot plot of a cell mixture consisting of 60% 2D6 clone and 40% line 33 co-cultured with the indicated melanoma is presented (right panel). (B) The percentage of CD107A+/CD8+ cells out of total CD8+ cells was correlated to the proportion of melanoma antigen-specific cells in the culture (R2=0.96, p<0.0001) (left panel). A representative dot plot of a cell mixture consisting of 60% 2D6 clone and 40% line 33 co-cultured with the indicated melanoma is shown (right panel) (C) The percentage of CSPG4+/CD8+ cells was correlated to the percentage of CD107A+/CD8+ cells (R2 = 0.98, p <0.0001) (left panel). A representative dot blot of a cell mixture consisting of 60% 2D6 clone and 40% line 33 co-cultured with the indicated melanoma is presented (right panel). One representative experiment of three is shown.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Lymphocyte imprinting with melanoma antigens acquired by trogocytosis facilitates identification of tumor-reactive T cells

doi: 10.4049/jimmunol.1202879

Figure Lengend Snippet: MART-126–35-specific T cells (clone 2D6) were mixed with melanoma non-relevant, MUC-163–71-specific T cells (line 33) at the indicated ratios, and co-cultured with HLA A2-matched melanoma cells (526mel) or irrelevant melanoma cells (888mel). (A) The percentage of CSPG4+/CD8+ melanoma antigen (MA)-imprinted cells out of total CD8+ cells was correlated to the proportion of melanoma antigen-specific cells in the culture (MA-specific cells, R2=0.99, p<0.0001) (left panel). A representative dot plot of a cell mixture consisting of 60% 2D6 clone and 40% line 33 co-cultured with the indicated melanoma is presented (right panel). (B) The percentage of CD107A+/CD8+ cells out of total CD8+ cells was correlated to the proportion of melanoma antigen-specific cells in the culture (R2=0.96, p<0.0001) (left panel). A representative dot plot of a cell mixture consisting of 60% 2D6 clone and 40% line 33 co-cultured with the indicated melanoma is shown (right panel) (C) The percentage of CSPG4+/CD8+ cells was correlated to the percentage of CD107A+/CD8+ cells (R2 = 0.98, p <0.0001) (left panel). A representative dot blot of a cell mixture consisting of 60% 2D6 clone and 40% line 33 co-cultured with the indicated melanoma is presented (right panel). One representative experiment of three is shown.

Techniques: Cell Culture, Dot Blot

TIL from three melanoma patients, M431, M412 and M209 (A, B and C respectively), were stimulated for 12 days with HLA-A2-matched melanoma cells and sorted to CSPG4-imprinted (CSPG4+/CD8+) and CSPG4-negative (CSPG4−/CD8+) by flow cytometry. Sorted cells were expanded in the presence of anti-CD3 antibody and co-cultured with HLA-A2-matched melanoma (526mel) or irrelevant (888mel) melanoma cells and analyzed for IFN-γ and TNF-α secretion by intracellular staining. The results of one out of three experiments are shown.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Lymphocyte imprinting with melanoma antigens acquired by trogocytosis facilitates identification of tumor-reactive T cells

doi: 10.4049/jimmunol.1202879

Figure Lengend Snippet: TIL from three melanoma patients, M431, M412 and M209 (A, B and C respectively), were stimulated for 12 days with HLA-A2-matched melanoma cells and sorted to CSPG4-imprinted (CSPG4+/CD8+) and CSPG4-negative (CSPG4−/CD8+) by flow cytometry. Sorted cells were expanded in the presence of anti-CD3 antibody and co-cultured with HLA-A2-matched melanoma (526mel) or irrelevant (888mel) melanoma cells and analyzed for IFN-γ and TNF-α secretion by intracellular staining. The results of one out of three experiments are shown.

Techniques: Flow Cytometry, Cell Culture, Staining

(A) TIL from melanoma patient 209 (M209) were sorted to gp100-positive and gp100-negative CD8+ T cells by flow cytometry. Sorted cells were expanded and co-cultured with HLA-A2-matched (624mel) or irrelevant (M171) melanoma cells. Percentage of specific lysis of target melanoma cells was measured using a 35S release-based cytotoxicity assay (p=0.016, left panel). TIL from M209 were sorted to CSPG4-imprinted (CSPG4+/CD8+) and CSPG4-negative (CSPG4−/CD8+) by flow cytometry. Cells were co-cultured with HLA-A2-matched (526mel, 624mel) or irrelevant (M171) melanoma cells. The level of intracellular cleaved caspase-3 in melanoma cells was measured by flow cytometry (right panel). (B) As for A, level of cleaved caspase-3 in melanomas after incubation with sorted TIL M431 or (C) with sorted TIL M412. The results of one out of three experiments are shown.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Lymphocyte imprinting with melanoma antigens acquired by trogocytosis facilitates identification of tumor-reactive T cells

doi: 10.4049/jimmunol.1202879

Figure Lengend Snippet: (A) TIL from melanoma patient 209 (M209) were sorted to gp100-positive and gp100-negative CD8+ T cells by flow cytometry. Sorted cells were expanded and co-cultured with HLA-A2-matched (624mel) or irrelevant (M171) melanoma cells. Percentage of specific lysis of target melanoma cells was measured using a 35S release-based cytotoxicity assay (p=0.016, left panel). TIL from M209 were sorted to CSPG4-imprinted (CSPG4+/CD8+) and CSPG4-negative (CSPG4−/CD8+) by flow cytometry. Cells were co-cultured with HLA-A2-matched (526mel, 624mel) or irrelevant (M171) melanoma cells. The level of intracellular cleaved caspase-3 in melanoma cells was measured by flow cytometry (right panel). (B) As for A, level of cleaved caspase-3 in melanomas after incubation with sorted TIL M431 or (C) with sorted TIL M412. The results of one out of three experiments are shown.

Techniques: Flow Cytometry, Cell Culture, Lysis, Cytotoxicity Assay, Incubation

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