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Western blot analysis of Olig2 on different lysates with Rabbit anti-Olig2 antibody (ET1604-29) at 1/5,000 dilution.
Lane 1: Mouse brain tissue lysate (20 µg/Lane)
Lane 2: Rat brain tissue lysate (20 µg/Lane)
Lane 3: Human brain tissue lysate (20 µg/Lane)
Predicted band size: 32 kDa
Observed band size: 36 kDa
Exposure time: 5 minutes 30 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1604-29) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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Fluorescence multiplex immunohistochemical analysis of mouse brain (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-NeuN (ET1602-12, red), anti-Iba1 (ET1705-78, green), anti-GFAP (ET1601-23, gray), anti-Olig2 (ET1604-29, cyan), anti-MAP2 (HA500177, magenta) and anti-CD34 (ET1606-11, yellow) on mouse brain. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in six rounds of staining: in the order of ET1602-12(1/5,000 dilution), ET1705-78 (1/2,000 dilution), ET1601-23 (1/5,000 dilution), ET1604-29 (1/1,000 dilution), HA500177 (1/5,000 dilution) and ET1606-11 (1/2,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
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Immunofluorescence analysis of frozen mouse cerebrum tissue with Rabbit anti-Myelin Basic Protein antibody (ET1702-15) at 1/500 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1702-15, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
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Western blot analysis of MAG on different lysates with Rabbit anti-MAG antibody (HA721818) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.
Lane 1: Mouse cerebellum tissue lysate
Lane 2: Mouse liver tissue lysate (negative)
Lane 3: Rat cerebellum tissue lysate
Lane 4: Rat liver tissue lysate (negative)
Lysates/proteins at 20 µg/Lane.
Predicted band size: 69 kDa
Observed band size: 100 kDa
Exposure time: 2 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721818) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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Western blot analysis of Myelin oligodendrocyte glycoprotein on different lysates with Rabbit anti-Myelin oligodendrocyte glycoprotein antibody (ET1705-16) at 1/1,000 dilution.
Lane 1: Mouse brain tissue lysate (40 µg/Lane)
Lane 2: Mouse lung tissue lysate (negative) (40 µg/Lane)
Lane 3: Rat brain tissue lysate (40 µg/Lane)
Lane 4: Rat lung tissue lysate (negative) (40 µg/Lane)
Predicted band size: 28 kDa
Observed band size: 28 kDa
Exposure time: 1 minute; ECL: K1802;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-16) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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