Eph receptor A2 Recombinant Rabbit Monoclonal Antibody [PSH09-21] - BSA and Azide free

This data was developed using HA723079, the same antibody clone in a different buffer formulation.
Western blot analysis of Eph receptor A2 on different lysates with Rabbit anti-Eph receptor A2 antibody (HA723079) at 1/2,000 dilution.

Lane 1: A549 cell lysate
Lane 2: HEK-293 cell lysate
Lane 3: HeLa cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 108 kDa
Observed band size: 130 kDa

Exposure time: 59 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 (HA723079) at 1/2,000 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.

This data was developed using HA723079, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of A549 cells labeling Eph receptor A2 with Rabbit anti-Eph receptor A2 antibody (HA723079) at 1/100 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-Eph receptor A2 antibody (HA723079) 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 (HA601187, 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 HA723079, the same antibody clone in a different buffer formulation.
Eph receptor A2 was immunoprecipitated from 0.2 mg A549 cell lysate with HA723079 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723079 at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: A549 cell lysate (input)
Lane 2: HA723079 IP in A549 cell lysate
Lane 3: Rabbit IgG instead of HA723079 in A549 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 59 seconds; ECL: K1801

This data was developed using HA723079, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of A549 cells labeling Eph receptor A2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA723079, 1/1,000) (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).