Rab3A Recombinant Rabbit Monoclonal Antibody [PSH01-08] - BSA and Azide free

☑ Relative expression (RE)

This data was developed using HA721573, the same antibody clone in a different buffer formulation.
Western blot analysis of Rab3A on different lysates with Rabbit anti-Rab3A antibody (HA721573) at 1/5,000 dilution.

Lane 1: Human brain tissue lysate
Lane 2: Human liver tissue lysate (negative)
Lane 3: Mouse brain tissue lysate
Lane 4: Mouse liver tissue lysate (negative)
Lane 5: Rat brain tissue lysate
Lane 6: Rat liver tissue lysate (negative)

Lysates/proteins at 40 µg/Lane.

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 27 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 (HA721573) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

☑ Knockdown (KD)

This data was developed using HA721573, the same antibody clone in a different buffer formulation.
Western blot analysis of Rab3A on different lysates with Rabbit anti-Rab3A antibody (HA721573) at 1/10,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Rab3A KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 1 minute; 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 (HA721573) at 1/10,000 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.

This data was developed using HA721573, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human cerebellum tissue with Rabbit anti-Rab3A antibody (HA721573) 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 (HA721573) 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.

This data was developed using HA721573, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-Rab3A antibody (HA721573) 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 (HA721573) 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.

This data was developed using HA721573, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-Rab3A antibody (HA721573) 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 (HA721573) 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.

This data was developed using HA721573, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of SH-SY5Y cells labeling Rab3A with Rabbit anti-Rab3A antibody (HA721573) at 1/100 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-Rab3A antibody (HA721573) at 1/100 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.

This data was developed using HA721573, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of N2A cells labeling Rab3A with Rabbit anti-Rab3A antibody (HA721573) at 1/100 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-Rab3A antibody (HA721573) at 1/100 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.

This data was developed using HA721573, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of PC-12 cells labeling Rab3A with Rabbit anti-Rab3A antibody (HA721573) at 1/100 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-Rab3A antibody (HA721573) at 1/100 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.

This data was developed using HA721573, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of N2A cells labeling Rab3A.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721573, 1ug/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).

This data was developed using HA721573, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of PC-12 cells labeling Rab3A.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721573, 1ug/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).