Olig2 Recombinant Rabbit Monoclonal Antibody [SP07-02]

Application: IHC-Fr

Species: Mouse

Site: Cerebral cortex

Sample: Frozen section

Antibody concentration: 1/500

Antigen retrieval: Not required

Application: IHC-Fr

Species: Mouse

Site: E14.5 embryo

Sample: Frozen section

Antibody concentration: 1/500

Antigen retrieval: The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.

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.

Fluorescence multiplex immunohistochemical analysis of mouse brain (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-MAP2 (HA500177, Red), anti-Olig2 (ET1604-29, Cyan), anti-GFAP (ET1601-23, Magenta) and anti-Neun (ET1602-12, 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 four rounds of staining: in the order of HA500177 (1/1,000 dilution), ET1604-29 (1/5,000 dilution), ET1601-23 (1/10,000 dilution) and ET1602-12 (1/10,000 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.

Fluorescence multiplex immunohistochemical analysis of mouse brain (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-Iba1 (ET1705-78, Green), anti-Olig2 (ET1604-29, White) and anti-TBR1 (ET1702-97, Red) on 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 three rounds of staining: in the order of ET1705-78 (1/2,000 dilution), ET1604-29 (1/1,000 dilution) and ET1702-97 (1/1,000 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 Zeiss Observer 7 Inverted Fluorescence Microscope.

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.

Immunohistochemical analysis of paraffin-embedded human glioma tissue with Rabbit anti-Olig2 antibody (ET1604-29) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1604-29) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-Olig2 antibody (ET1604-29) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1604-29) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Olig2 antibody (ET1604-29) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1604-29) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Olig2 antibody (ET1604-29) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1604-29) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Application: IF-tissue

Species: Mouse

Site: Hippocampus

Sample: Paraffin-embedded section

Antibody concentration: 1/200

Immunocytochemistry analysis of mouse glial cells labeling Olig2 with Rabbit anti-Olig2 antibody (ET1604-29) at 1/200 dilution.

Cells were fixed with 4% PFA (15 min), permeabilized with 0.25% TritonX-100 for 15 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4℃ with Rabbit anti-Olig2 antibody (ET1604-29) at at 1/200 dilution. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

☑ Relative expression (RE)

Western blot analysis of Olig2 on different lysates with Rabbit anti-Olig2 antibody (ET1604-29) at 1/5,000 dilution.

Lane 1: Mouse brain (P0) tissue lysate
Lane 2: Mouse brain tissue lysate
Lane 3: Mouse liver tissue lysate (negative)
Lane 4: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 32 kDa
Observed band size: 36 kDa

Exposure time: 42 seconds; 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 (ET1604-29) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.