35-6200

antibody from Invitrogen Antibodies

Targeting: CTAG1A ESO1, LAGE2A

Provider product page for 35-6200

Western blot

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [38]
Comments [0]
Validations
Immunohistochemistry [1]
Other assay [19]

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Product number: 35-6200 - Provider product page
Provider: Invitrogen Antibodies
Product name: NY-ESO-1 Monoclonal Antibody (E978)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: This antibody can react specifically with the 23 kDa NY-ESO-1 protein. During development reactivity was confirmed with human SKMEL-37 melanoma cells.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: E978
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

CTAG1A protein structure - 35-6200 shown in red.

Supportive validation

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Experimental details: NULL

Supportive validation

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Experimental details: NULL

Supportive validation

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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

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Experimental details: NULL

Supportive validation

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Experimental details: Figure 2 Comparison of GAGE, MAGE-1 and NY-ESO-1 staining in normal testis, and analysis of GAGE protein expression in ovary and different types of cancer. GAGE ( A ), MAGE ( B ) and NY-ESO-1 ( C ) were detected in spermatogonia and primary spermatocytes of the seminiferous tubuli. However, while MAGE-A1 and NY-ESO-1 were located only in the cytoplasm of these cells, GAGE staining was also present, and more intense, in the nuclei. GAGE was also expressed in oocytes of resting ( D ) and maturing ( E ) follicles of normal ovaries, which also contained GAGE-negative oocytes. In three malignant melanomas, all cells exhibited cytoplasmic staining ( G - I ), whereas nuclear staining was observed in only two melanomas ( G and H ). Heterogeneous staining was also seen in other types of cancer, including lung carcinoma ( F ) and breast carcinoma ( J - L ). One breast carcinoma exhibited variations in both cytoplasmic and nuclear GAGE staining among cells ( J and K ), while only few cells of another breast cancer specimen were positive ( L ). Magnification: x 10 ( A , B , C , E , F , J , L ), x 20 ( G - I ), x 40 ( D , K ).

Supportive validation

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Experimental details: Figure 3 NY-ESO-1 serum autoantibodies and antigen expression in high-grade serous ovarian cancer. (A) Serum autoantibody responses to NY-ESO-1 in the 35-patient cohort. Autoantibody responses are reported as the number of standard deviations from the mean of 60 age- and gender-matched controls with no known personal history of cancer. (B,C) Immunohistochemical analysis of NY-ESO-1 expression in two representative serous ovarian tumors with high (B) and negative (C) expression of the antigen.

Supportive validation

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Experimental details: Figure 4 Analysis of the T cell response in a patient with autoantibodies to NY-ESO-1 showing the presence of NY-ESO-1 reactive CD8+ T cells in ascites and tumor despite the lack of NY-ESO-1 expression in solid tumor. (A) MHC class I pentamer analysis demonstrating enrichment of NY-ESO-1-specific CD8+ T cells in ascites and solid tumor compared to peripheral blood. The boxed areas and associated numbers represent the percentage of pentamer-positive cells relative to total CD8+ cells. (B) ELISPOT analysis of IFN-gamma production by T cells after stimulation with an HLA-A2-binding peptide from NY-ESO-1. Data is presented as the number of IFN-gamma-producing cells per 1x10 6 bulk cells from the indicated tissue compartments. (C) Immunohistochemical analysis of tumor-infiltrating CD3+ and CD8+ T cells in tumor stroma, and expression of NY-ESO-1 antigen. While the solid tumor was negative for expression of NY-ESO-1, a fraction of cells from ascites were positive. The cellular fraction of ascites also contained cytokeratin-positive epithelial cells, presumably of tumor origin (data not shown).

Supportive validation

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Experimental details: Figure 2 Representative immunohistochemical expression patterns from 1-10%, 11-50% and 51-100% of MAGE-C2/CT10 (A, B and C), MAGE-C1/CT7 (D, E and F) and NY-ESO-1 (G, H and I). A: Radical prostatectomy specimen (Gleason 4+3), B: Bone metastasis of prostate cancer, C: Bone metastasis of prostate cancer, D: Radical prostatectomy specimen (Gleason 4+3), E: Castration-resistant prostate cancer (Gleason 5+5), F: Palliative transurethral resection of prostate cancer (Gleason 4+5), G: Bone metastasis of prostate cancer, H: Palliative transurethral resection of prostate cancer (Gleason 5+4), I: Castration-resistant prostate cancer (Gleason 5+4). Original magnification: 200x; Magnification bar: 20 uM.

Supportive validation

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Experimental details: Figure 2 gamma-radiation up-regulates CT-antigens and MHC-I molecules on the protein level. (A) Immunofluorescence of MDA-MB-469 and SK-MEL-37 cells exposed to a single dose irradiation of 20 Gy and stained with antibodies against NY-ESO-1 and CT7 at different time points after irradiation. (B) MDA-MB-469, SK-MEL-37 and MCF 10A cell lines were exposed to a single dose of 20 Gy and the expression of NY-ESO-1 and CT7 was analyzed by Western blotting 72 h later. (C) breast cancer and (D) osteosarcoma cell lines were exposed to single dose irradiation of 20 Gy and the expression of HLA-ABC and beta 2 microglobulin was quantified at different time points after irradiation by flow cytometry. RAD: indicates gamma-radiation.

Supportive validation

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Experimental details: Figure 5 Radiotherapy induced expression of CT-antigens and MHC-I and lymphocyte infiltration in sarcoma patients. Paraffin-embedded paired tissue sections obtained from sarcoma patients (n = 15) before and after irradiation were analyzed by immunohistochemistry for (A) presence of CD8 + , CD4 + and granzyme + cells, (B) expression of CT7, NY-ESO-1 and CT10 and (C) MHC-I expression. The CT-antigens were scored as the mean percentage of live cells expressing the antigen to the total number of cells in five high power fields (40X objective). The infiltration by lymphocytes was taken as the mean by counting the number of cells expressing CD4, CD8 and granzyme in five high power fields, and MHC-I was scored based on the intensity of the staining. (D) Representative sections showing the expression of CT-antigens and MHC-I and infiltration of lymphocytes before and after radiotherapy by immunohistochemistry. Depicted are: patient F for the expression of CT7, patient A for CT10, patient K for NY-ESO-1, patient D for CD4 and MHC-I and patient E for CD8 expression. NR indicates non-radiated and RAD indicates corresponding irradiated sections. Pt: indicates patient number. Patient information is listed in supplementary Table S2 .

Supportive validation

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Experimental details: Fig 2 Effects of DAC treatment on NY-ESO-1 mRNA and protein expression in MCF7, U266, and ARK cell lines. A. NY-ESO-1 specific mRNA, quantified as copy numbers/mug RNA using qRT-PCR. All data are representative of three independent experiments performed in triplicate. B. NY-ESO-1 protein expression analyzed by Western blotting (n = 3). The first and second line show total cell lysate of the respective cell line +/- DAC treatment and detection via a NY-ESO-1 specific or alpha-Tubulin specific (loading control) antibody. The third line shows a Western blot of recombinant NY-ESO-1 protein as control.

Supportive validation

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Experimental details: Figure 2 Effect of radiation on cell surface NY-ESO-1 measurement by fluorescence flow cytometry. Sub-confluent adherent cultures of indicated cell lines that had been grown in parallel to same cell density were treated with 0 or 15 Gy X-rays. After four days, adherent cells were collected by scraping and examined by flow cytometry for binding of an unconjugated mouse IgG1 antibody against NY-ESO-1. Antibody binding was indirectly detected with a fluorophore-conjugated rat antibody against mouse IgG. Binding of a control IgG1 (normal mouse serum fraction) was similarly detected. Shown are representative histograms of viable cells identified by 7 -amino-actinomycin D staining.

Supportive validation

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Experimental details: Figure 1. Immunohistochemical findings for MAGE-A4, NY-ESO-1 and CD33 in non-small cell lung cancer tissues. Specimens from the normal lung were used as a negative control. Staining of the normal lung demonstrated no reactivity for (A) CD33, (B) MAGE-A4 and (C) NY-ESO-1. Expression of (D) CD33, (E) MAGE-A4 and (F) NY-ESO-1 in squamous cell carcinoma. Expression of (G) CD33, (H) MAGE-A4 and (I) NY-ESO-1 in adenocarcinoma. Magnification, x100. MDSCs, myeloid derived suppressor cells; MAGE-A4, melanoma-associated antigen 4; NY-ESO-1, New York esophageal squamous cell carcinoma-1.

Supportive validation

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Experimental details: Figure 1 Representative images of immunohistochemistry for B7-H3, B7-H4, PD-L1, PD-L2, MAGE-A, NY-ESO-1, MHC-I, and B2M in in situ and invasive EMPD. IHC images for each marker along with the corresponding H&E image for each case are shown for in situ and invasive EMPD.