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mouse anti mage a3 monoclonal antibody (Novus Biologicals)

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Novus Biologicals mouse anti mage a3 monoclonal antibody

( a ) K562 aAPCs were transiently transfected with the indicated combinations of genes and cultured in the presence or absence of 0.02 μM bortezomib or 0.02 μM carfilzomib for 48 h. Total cell lysates were immunoblotted with <t>anti-MAGE-A3,</t> anti-HLA-class I or anti-β-actin <t>mAb.</t> ( b , c ) K562/DP4/Ii cells were transiently transfected with a retrovirus vector encoding IRES-EGFP (control), or a native ( b ) or endosome-targeted ( c ) form of MAGE-A3 linked with IRES-EGFP. Cells were then cultured with carfilzomib at the indicated concentrations for 48 h. Transient transfection efficiencies were normalized to EGFP expression measured by flow cytometry. DP4/MAGE-A3 243–258 CD4 + T cells were stimulated with the K562/DP4/Ii transfectants and IFN-γ secretion was measured by ELISPOT analysis. Data shown represent means±s.d.'s of triplicates. ( d ) DP4/MAGE-A3 243–258 CD4 + T cells were stimulated with the indicated K562-based aAPCs pulsed with tetanus toxin 947–967 (control) or MAGE-A3 243–258 peptide, and IFN-γ secretion was evaluated by ELISPOT assays. Data shown represent means±s.d.'s of triplicates. ( e , f ) The indicated K562-based aAPCs were transiently transfected with a retrovirus vector encoding IRES-EGFP (control) or a native ( e ) or endosome-targeted ( f ) form of MAGE-A3 linked with IRES-EGFP. Transient transfection efficiencies were normalized to EGFP expression measured by flow cytometry. DP4/MAGE-A3 243–258 CD4 + T cells were stimulated with the indicated aAPCs and IFN-γ secretion was measured by ELISPOT analysis. Data shown represent means±s.d.'s of triplicates. Results are representative of three independent experiments. ns, not significant; * P <0.05 by unpaired, two-tailed Welch's t -test.

Mouse Anti Mage A3 Monoclonal Antibody, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+mage+a3+monoclonal+antibody/MAGEA3+Antibody+(4D9)/pmc05436232-187-65-71
Average 90 stars, based on 1 article reviews

mouse anti mage a3 monoclonal antibody - by Bioz Stars, 2026-09

90/100 stars

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1) Product Images from "HLA-DP 84Gly constitutively presents endogenous peptides generated by the class I antigen processing pathway"

Article Title: HLA-DP 84Gly constitutively presents endogenous peptides generated by the class I antigen processing pathway

Journal: Nature Communications

doi: 10.1038/ncomms15244

Figure Legend Snippet: ( a ) K562 aAPCs were transiently transfected with the indicated combinations of genes and cultured in the presence or absence of 0.02 μM bortezomib or 0.02 μM carfilzomib for 48 h. Total cell lysates were immunoblotted with anti-MAGE-A3, anti-HLA-class I or anti-β-actin mAb. ( b , c ) K562/DP4/Ii cells were transiently transfected with a retrovirus vector encoding IRES-EGFP (control), or a native ( b ) or endosome-targeted ( c ) form of MAGE-A3 linked with IRES-EGFP. Cells were then cultured with carfilzomib at the indicated concentrations for 48 h. Transient transfection efficiencies were normalized to EGFP expression measured by flow cytometry. DP4/MAGE-A3 243–258 CD4 + T cells were stimulated with the K562/DP4/Ii transfectants and IFN-γ secretion was measured by ELISPOT analysis. Data shown represent means±s.d.'s of triplicates. ( d ) DP4/MAGE-A3 243–258 CD4 + T cells were stimulated with the indicated K562-based aAPCs pulsed with tetanus toxin 947–967 (control) or MAGE-A3 243–258 peptide, and IFN-γ secretion was evaluated by ELISPOT assays. Data shown represent means±s.d.'s of triplicates. ( e , f ) The indicated K562-based aAPCs were transiently transfected with a retrovirus vector encoding IRES-EGFP (control) or a native ( e ) or endosome-targeted ( f ) form of MAGE-A3 linked with IRES-EGFP. Transient transfection efficiencies were normalized to EGFP expression measured by flow cytometry. DP4/MAGE-A3 243–258 CD4 + T cells were stimulated with the indicated aAPCs and IFN-γ secretion was measured by ELISPOT analysis. Data shown represent means±s.d.'s of triplicates. Results are representative of three independent experiments. ns, not significant; * P <0.05 by unpaired, two-tailed Welch's t -test.

Techniques Used: Transfection, Cell Culture, Plasmid Preparation, Control, Expressing, Flow Cytometry, Enzyme-linked Immunospot, Two Tailed Test

( a , b ) The indicated K562-based aAPCs were transiently transfected with a retrovirus vector encoding IRES-EGFP (control) or a native ( a ) or endosome-targeted ( b ) form of MAGE-A3 linked with IRES-EGFP. Transient transfection efficiencies were normalized to EGFP expression measured by flow cytometry. DP4/MAGE-A3 243–258 CD4 + T cells were stimulated with the indicated APCs and IL-2 secretion was measured by ELISPOT analysis. Data shown represent means±s.d.'s of triplicates. ( c ) NSG mice were subcutaneously inoculated with 2 × 10 6 K562 cells stably expressing DP4/Ii/MAGE-A3 or DP4 84DEAV87 /Ii/MAGE-A3. Two days later, the mice were treated with 4 × 10 7 CD3 + T cells untransduced or transduced with DP4/MAGE-A3 243–258 TCR. The mean tumour size for each group is represented as the average±s.d. of three mice. There was no significant difference in the tumorigenicity of the two cell lines (data not shown). Results are representative of three independent experiments. ns, not significant; * P <0.05, ** P <0.01 by unpaired, two-tailed Welch's t -test.

Figure Legend Snippet: ( a , b ) The indicated K562-based aAPCs were transiently transfected with a retrovirus vector encoding IRES-EGFP (control) or a native ( a ) or endosome-targeted ( b ) form of MAGE-A3 linked with IRES-EGFP. Transient transfection efficiencies were normalized to EGFP expression measured by flow cytometry. DP4/MAGE-A3 243–258 CD4 + T cells were stimulated with the indicated APCs and IL-2 secretion was measured by ELISPOT analysis. Data shown represent means±s.d.'s of triplicates. ( c ) NSG mice were subcutaneously inoculated with 2 × 10 6 K562 cells stably expressing DP4/Ii/MAGE-A3 or DP4 84DEAV87 /Ii/MAGE-A3. Two days later, the mice were treated with 4 × 10 7 CD3 + T cells untransduced or transduced with DP4/MAGE-A3 243–258 TCR. The mean tumour size for each group is represented as the average±s.d. of three mice. There was no significant difference in the tumorigenicity of the two cell lines (data not shown). Results are representative of three independent experiments. ns, not significant; * P <0.05, ** P <0.01 by unpaired, two-tailed Welch's t -test.

Techniques Used: Transfection, Plasmid Preparation, Control, Expressing, Flow Cytometry, Enzyme-linked Immunospot, Stable Transfection, Transduction, Two Tailed Test

Western Blot:

Article Title: HLA-DP 84Gly constitutively presents endogenous peptides generated by the class I antigen processing pathway
Article Snippet: After blocking with 0.1% Tween 20 in Tris-buffered saline, the membranes were incubated with the indicated primary antibodies at 4 °C overnight, washed and incubated with horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (H+L) secondary antibody (Promega, Madison, WI) or HRP-conjugated rat anti-mouse IgG VeriBlot secondary antibody (Abcam, Cambridge, MA), which is only capable of recognizing native, non-denatured primary mouse antibody, at room temperature for 1 h. The signal was detected by enhanced chemiluminescence (GE Healthcare). .. Antibodies to the following proteins were used in immunoblots: mouse anti-Ii monoclonal antibodies (sc-6262, 1:1,000, and sc-47742, 1:500, Santa Cruz Biotechnology), mouse anti-DR/DP monoclonal antibody (sc-51617, 1:1,000, Santa Cruz Biotechnology), mouse anti-DR/DP monoclonal antibody (sc-51617, 1:2,000, Santa Cruz Biotechnology), mouse anti-DP monoclonal antibody (H127, 1:1,000, Leinco Technologies), mouse anti-DRα monoclonal antibody (sc-53499, 1:1,000, Santa Cruz Biotechnology), mouse anti-CLIP monoclonal antibody (sc-12725, 1:3,000, Santa Cruz Biotechnology), mouse anti-MAGE-A3 monoclonal antibody (H00004102-M0, 1:1,000, Novus Biologicals) and mouse anti-HLA class I monoclonal antibody (ab70328, 1:2,000, Abcam). ..

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Journal: Nature Communications

Article Title: HLA-DP 84Gly constitutively presents endogenous peptides generated by the class I antigen processing pathway

doi: 10.1038/ncomms15244

Figure Lengend Snippet: ( a ) K562 aAPCs were transiently transfected with the indicated combinations of genes and cultured in the presence or absence of 0.02 μM bortezomib or 0.02 μM carfilzomib for 48 h. Total cell lysates were immunoblotted with anti-MAGE-A3, anti-HLA-class I or anti-β-actin mAb. ( b , c ) K562/DP4/Ii cells were transiently transfected with a retrovirus vector encoding IRES-EGFP (control), or a native ( b ) or endosome-targeted ( c ) form of MAGE-A3 linked with IRES-EGFP. Cells were then cultured with carfilzomib at the indicated concentrations for 48 h. Transient transfection efficiencies were normalized to EGFP expression measured by flow cytometry. DP4/MAGE-A3 243–258 CD4 + T cells were stimulated with the K562/DP4/Ii transfectants and IFN-γ secretion was measured by ELISPOT analysis. Data shown represent means±s.d.'s of triplicates. ( d ) DP4/MAGE-A3 243–258 CD4 + T cells were stimulated with the indicated K562-based aAPCs pulsed with tetanus toxin 947–967 (control) or MAGE-A3 243–258 peptide, and IFN-γ secretion was evaluated by ELISPOT assays. Data shown represent means±s.d.'s of triplicates. ( e , f ) The indicated K562-based aAPCs were transiently transfected with a retrovirus vector encoding IRES-EGFP (control) or a native ( e ) or endosome-targeted ( f ) form of MAGE-A3 linked with IRES-EGFP. Transient transfection efficiencies were normalized to EGFP expression measured by flow cytometry. DP4/MAGE-A3 243–258 CD4 + T cells were stimulated with the indicated aAPCs and IFN-γ secretion was measured by ELISPOT analysis. Data shown represent means±s.d.'s of triplicates. Results are representative of three independent experiments. ns, not significant; * P <0.05 by unpaired, two-tailed Welch's t -test.

Article Snippet: Antibodies to the following proteins were used in immunoblots: mouse anti-Ii monoclonal antibodies (sc-6262, 1:1,000, and sc-47742, 1:500, Santa Cruz Biotechnology), mouse anti-DR/DP monoclonal antibody (sc-51617, 1:1,000, Santa Cruz Biotechnology), mouse anti-DR/DP monoclonal antibody (sc-51617, 1:2,000, Santa Cruz Biotechnology), mouse anti-DP monoclonal antibody (H127, 1:1,000, Leinco Technologies), mouse anti-DRα monoclonal antibody (sc-53499, 1:1,000, Santa Cruz Biotechnology), mouse anti-CLIP monoclonal antibody (sc-12725, 1:3,000, Santa Cruz Biotechnology), mouse anti-MAGE-A3 monoclonal antibody (H00004102-M0, 1:1,000, Novus Biologicals) and mouse anti-HLA class I monoclonal antibody (ab70328, 1:2,000, Abcam).

Techniques: Transfection, Cell Culture, Plasmid Preparation, Control, Expressing, Flow Cytometry, Enzyme-linked Immunospot, Two Tailed Test

Journal: Nature Communications

Article Title: HLA-DP 84Gly constitutively presents endogenous peptides generated by the class I antigen processing pathway

doi: 10.1038/ncomms15244

Figure Lengend Snippet: ( a , b ) The indicated K562-based aAPCs were transiently transfected with a retrovirus vector encoding IRES-EGFP (control) or a native ( a ) or endosome-targeted ( b ) form of MAGE-A3 linked with IRES-EGFP. Transient transfection efficiencies were normalized to EGFP expression measured by flow cytometry. DP4/MAGE-A3 243–258 CD4 + T cells were stimulated with the indicated APCs and IL-2 secretion was measured by ELISPOT analysis. Data shown represent means±s.d.'s of triplicates. ( c ) NSG mice were subcutaneously inoculated with 2 × 10 6 K562 cells stably expressing DP4/Ii/MAGE-A3 or DP4 84DEAV87 /Ii/MAGE-A3. Two days later, the mice were treated with 4 × 10 7 CD3 + T cells untransduced or transduced with DP4/MAGE-A3 243–258 TCR. The mean tumour size for each group is represented as the average±s.d. of three mice. There was no significant difference in the tumorigenicity of the two cell lines (data not shown). Results are representative of three independent experiments. ns, not significant; * P <0.05, ** P <0.01 by unpaired, two-tailed Welch's t -test.

Techniques: Transfection, Plasmid Preparation, Control, Expressing, Flow Cytometry, Enzyme-linked Immunospot, Stable Transfection, Transduction, Two Tailed Test

Figure 1. Prediction and validation of HCA587/MAGE-C2 as the target of miR-874. (A) Quantitative RT-PCR analysis of the relative HCA587 mRNA expression in A375 cells transfected with each mimic of 8 candidate miRNAs for 48 h. (B) Western blot analysis of the endogenous HCA587 protein level in A375 cells transfected with each mimic of 8 candidate miRNAs for 48 h. The quantitative analysis is shown in the right panel. (C) Western blot analysis of the endogenous HCA587, MAGE-A1 and MAGE-A3 protein levels in A375 cells transfected with miR-874 mimic for 48 h. (D) Schematic of putative binding site of miR-874 in HCA587 3’-UTR. A mutant HCA587 3’-UTR construct (position 34-40 deletion) was tested in parallel. (E) Analysis of luciferase activity. Renilla luciferase reporters containing either the wild-type or mutant form of HCA587 3’-UTR were co-transfected into HEK293T cells with miR-874 mimic or miR-NC. At 48 h post-transfection, Renilla luciferase activity was measured. The results were normalized against firefly luciferase values. (mean ± S.D.; *** p < 0.001).

Journal: Journal of Cancer

Article Title: MicroRNA-874 Functions as a Tumor Suppressor by Targeting Cancer/Testis Antigen HCA587/MAGE-C2.

doi: 10.7150/jca.13674

Figure Lengend Snippet: Figure 1. Prediction and validation of HCA587/MAGE-C2 as the target of miR-874. (A) Quantitative RT-PCR analysis of the relative HCA587 mRNA expression in A375 cells transfected with each mimic of 8 candidate miRNAs for 48 h. (B) Western blot analysis of the endogenous HCA587 protein level in A375 cells transfected with each mimic of 8 candidate miRNAs for 48 h. The quantitative analysis is shown in the right panel. (C) Western blot analysis of the endogenous HCA587, MAGE-A1 and MAGE-A3 protein levels in A375 cells transfected with miR-874 mimic for 48 h. (D) Schematic of putative binding site of miR-874 in HCA587 3’-UTR. A mutant HCA587 3’-UTR construct (position 34-40 deletion) was tested in parallel. (E) Analysis of luciferase activity. Renilla luciferase reporters containing either the wild-type or mutant form of HCA587 3’-UTR were co-transfected into HEK293T cells with miR-874 mimic or miR-NC. At 48 h post-transfection, Renilla luciferase activity was measured. The results were normalized against firefly luciferase values. (mean ± S.D.; *** p < 0.001).

Article Snippet: After blocking, the membrane was incubated with specific testing antibodies for HCA587/MAGE-C2, MAGE-A1 (Bioworld, USA), or MAGE-A3 (Origene, USA) and GAPDH (Bioworld).

Techniques: Biomarker Discovery, Quantitative RT-PCR, Expressing, Transfection, Western Blot, Binding Assay, Mutagenesis, Construct, Luciferase, Activity Assay

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mouse anti mage a3 monoclonal antibody (Novus Biologicals)

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