GFP Recombinant Rabbit Monoclonal Antibody [SP00-68]

Western blot analysis of GFP on different lysates with Rabbit anti-GFP antibody (ET1604-25) at 1/20,000 dilution.

Lane 1: 293T cell lysate
Lane 2: 293T transfected with GFP cell lysate

Lysates/proteins at 5 µg/Lane.

Predicted band size: 27 kDa
Observed band size: 27 kDa

Exposure time: 2 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 (ET1604-25) at 1/20,000 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.

Immunocytochemistry analysis of HeLa cells transfected with GFP labeling GFP with Rabbit anti-GFP antibody (ET1604-25, red) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-GFP antibody (ET1604-25, red) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) 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.

GFP tag was immunoprecipitated in 5µg GFP Tag fusion protein lysate with ET1604-25 at 2 µg/20 µl agarose. Western blot was performed from the immunoprecipitate using M1004-8 at 1/1000 dilution. Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 60 mins at room temperature.

Lane 1: GFP Tag fusion protein lysate (input).
Lane 2: Rabbit IgG instead of ET1604-25 in GFP Tag fusion protein lysate.
Lane 3: ET1602-7 IP in GFP Tag fusion protein lysate.
Lane 4: ET1604-25 IP in GFP Tag fusion protein lysate.
Lane 5: ET1604-26 IP in GFP Tag fusion protein lysate.
Lane 6: ET1607-31 IP in GFP Tag fusion protein lysate.
Lane 7: R1312-2 IP in GFP Tag fusion protein lysate.

Blocking/Dilution buffer: 5% NFDM/TBST

Immunohistochemical analysis of paraffin-embedded mouse brain tissue transfected with GFP-tagged CX3CR1 with Rabbit anti-GFP antibody (ET1604-25) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1604-25) 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 transfected with GFP-tagged CX3CR1 with Rabbit anti-GFP antibody (ET1604-25) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1604-25) 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.