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Figure 4 Immunocytochemistry Validation of BAFF in HL60 CellsImmunocytochemical analysis of HL60 cells using anti-BAFF antibody (2221) at 1 &956,g/ml. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT, antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4&730,C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. |
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Figure 5 Immunofluorescence Validation of BAFF in Human Spleen TissueImmunofluorescent analysis of 4% paraformaldehyde-fixed human spleen tissue labeling BAFF with 2221 at 20 &956,g/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue). |
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Figure 7 Immunohistochemistry Validation of BAFF in Thyroid of Patients with Graves Diseases (Campi et al., 2015) BAFF expression detected by anti-BAFF antibodies (2221) was remarkably increased in thyrocytes from multinodular goiter (C) compared with either Hashimotos thyroiditis (E) or Graves disease (G) while no staining was found in normal thyroid tissue (A). |
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Figure 8 Immunohistochemistry Validation of BAFF in Murine Cardiac Transplants at Rejection (Ye et al., 2004) BAFF expression detected by anti-BAFF antibodies (2221) was upregulated in intragraft leukocytes due to rejection at 7 days after heart transplant. |
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Figure 1 Western Blot Validation in Human HL60 Cell Lysate (H) and Mouse Spleen Lysate (M) Loading: 15 &956,g of lysates per lane.Antibodies: BAFF 2221 (1 &956,g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |
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Figure 2 Western Blot Validation in Human, Mouse and Rat Cell LinesLoading: 15 &956,g of lysates per lane.Antibodies: BAFF 2221 (1 &956,g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |
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Figure 3 Western Blot Validation with Recombinant ProteinLoading: 30 ng of human BAFF recombinant protein per lane.Antibodies: BAFF 2221 (Lane 1: 0.25 &956,g/mL, Lane 2: 0.5 &956,g/mL and Lane 3: 1 &956,g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Observed at around 18kD. |
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Figure 6 Regulated Expression Validation of BAFF in Myeloma Patients (Tai et al., 2006) Immunoblot analysis was performed to monitor protein expression of BAFF with anti-BAFF antibodies in multiple myeloma cells with or without BMSCs. BAFF expression in cocultures at 8hr or 24hr was up-regulated by ~3.5-fold relative to BMSCs alone. |