Formalin-fixed paraffin-embedded tumor sections were immunohistochemically stained with mouse anti-human GD2 antibody (BD Biosciences, Cat# 554,272, 0.5 mg/mL). The isotype control was stained with purified mouse IgG2a (BioLegend, Cat# 400,202). All images were captured from tumor sections using a Zeiss Axio Vert.A1 microscope and Zeiss ZEN imaging software. The tissue staining intensity was compared with that from positive and negative controls and scored from 0 to 4 according to two components: staining intensity and the percentage of positive cells. Each sample was assessed and graded by two independent observers.
Mouse anti human gd2 antibody
Manufactured by BD
The mouse anti-human GD2 antibody is a laboratory reagent used to detect and quantify the expression of the GD2 ganglioside on cell surfaces. GD2 is a disialoganglioside that is commonly expressed on the surface of various cell types, including neurons and certain types of cancer cells. The antibody can be used in techniques such as flow cytometry, immunohistochemistry, and enzyme-linked immunosorbent assay (ELISA) to measure the presence and levels of GD2 in samples.
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2 protocols using mouse anti human gd2 antibody
1
Quantitative GD2 Expression Analysis
2
Immunohistochemical Analysis of Neuronal Markers
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Immunohistochemical (IHC) analysis was performed on formalin-fixed, paraffin-embedded (FFPE) tissues as previously described (34) using rabbit-polyclonal anti-CHD5 antibody (Strategic Diagnostics, DE, USA) at a 1:1000 dilution for 1 h; mouse-polyclonal anti-neurofilament protein, 68 kD (NF68) antibody (Invitrogen. Waltham, Massachusetts. USA) 1:300 dilution, 1 h; mouse-polyclonal anti-Glial fibrillary acidic protein (GFAP) antibody (Novocastra, UK) 1:200 dilution, 2 min; rabbit-polyclonal S-100 protein antibody (Diagnostic Biosystems. Pleasanton, California. USA) 1:800 dilution, 15 min. GD2 analysis was performed by IHC analysis on frozen tissues using mouse anti-human GD2 antibody (BD Pharmingen. San Diego, California. USA) 1:300 dilution, 15 min.
All slides were examined by pediatric pathologist (M.S.) using an Olympus BX41 light microscopy. Assessing staining and score of both percentage of positive cells and staining intensity were done as follows: 0, negative; 1, weak; 2, strong; and 3, very intense staining. Integer values were assigned to the proportion of positive cells: 25% = 1; 25–75% = 2; >75% = 3. For CHD5, intensity and positive cell values were multiplied to provide a single score for each case.
All slides were examined by pediatric pathologist (M.S.) using an Olympus BX41 light microscopy. Assessing staining and score of both percentage of positive cells and staining intensity were done as follows: 0, negative; 1, weak; 2, strong; and 3, very intense staining. Integer values were assigned to the proportion of positive cells: 25% = 1; 25–75% = 2; >75% = 3. For CHD5, intensity and positive cell values were multiplied to provide a single score for each case.
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