Otx2 Recombinant Rabbit Monoclonal Antibody [PSH01-61] - BSA and Azide free

Application: IHC-Fr

Species: Mouse

Site: E14.5 embryo

Sample: Frozen section

Antibody concentration: 1:500

Antigen retrieval: Not required

Immunohistochemical analysis of paraffin-embedded mouse embryo tissue with Rabbit anti-Otx2 antibody (HA750737) 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 (HA750737) 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 embryo tissue with Rabbit anti-Otx2 antibody (HA750737) 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 (HA750737) 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 eye tissue with Rabbit anti-Otx2 antibody (HA750737) 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 (HA750737) 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 eye tissue with Rabbit anti-Otx2 antibody (HA750737) at 1/200 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 (HA750737) at 1/200 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.

☑ Cell treatment (CT)

Western blot analysis of Otx2 on different lysates with Rabbit anti-Otx2 antibody (HA750737) at 1/1,000 dilution.

Lane 1: 293T transfected with FLAG-tagged empty control cell lysate
Lane 2: 293T transfected with FLAG-tagged Otx2 cell lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 32 kDa
Observed band size: 35 kDa

Exposure time: 24 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 (HA750737) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

Western blot analysis of Otx2 on different lysates with Rabbit anti-Otx2 antibody (HA750737) at 1/1,000 dilution.

Lane 1: Mouse eyeball tissue lysate (40 µg/Lane)
Lane 2: Rat embryo tissue lysate (40 µg/Lane)
Lane 3: Rat eyeball tissue lysate (40 µg/Lane)

Predicted band size: 32 kDa
Observed band size: 35 kDa

Exposure time: 5 minutes 10 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 (HA750737) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Western blot analysis of Otx2 on Y79 cell lysates with Rabbit anti-Otx2 antibody (HA750737) at 1/5,000 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 32 kDa
Observed band size: 35 kDa

Exposure time: 9 seconds; 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 (HA750737) 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.

Application: IF-tissue

Species: Mouse

Site: E14.5 embryo

Sample: Paraffin-embedded section

Antibody concentration: 1:500

☑ Cell treatment (CT)

Immunocytochemistry analysis of 293T overexpress with or without Otx2 cells labeling Otx2 with Rabbit anti-Otx2 antibody (HA750737) at 1/10,000 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Otx2 antibody (HA750737) at 1/10,000 dilution in 1% BSA in PBST overnight at 4 ℃. 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.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

☑ Cell treatment (CT)

Flow cytometric analysis of 293T overexpress with or without Otx2 cells labeling Otx2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA750737, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).