MAPK6 / ERK3 Recombinant Rabbit Monoclonal Antibody [PSH03-39] - BSA and Azide free

This data was developed using HA721984, the same antibody clone in a different buffer formulation.
Western blot analysis of MAPK6 / ERK3 on different lysates with Rabbit anti-MAPK6 / ERK3 antibody (HA721984) at 1/2,000 dilution.

Lane 1: A431 cell lysate
Lane 2: A549 cell lysate
Lane 3: PC-3M cell lysate
Lane 4: C2C12 cell lysate
Lane 5: NIH/3T3 cell lysate
Lane 6: C6 cell lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 5 minutes; ECL: K1801


Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA721984, 1/2,000 in 5% NFDM/TBST, overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 83 kDa
Observed band size: 105 kDa

☑ Knockdown (KD)

This data was developed using HA721984, the same antibody clone in a different buffer formulation.
Western blot analysis of MAPK6 / ERK3 on different lysates with Rabbit anti-MAPK6 / ERK3 antibody (HA721984) at 1/5,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-MAPK6 / ERK3 KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 83 kDa
Observed band size: 100 kDa

Exposure time: 40 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 (HA721984) 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.

This data was developed using HA721984, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of 293T cells labeling MAPK6 / ERK3 with Rabbit anti-MAPK6 / ERK3 antibody (HA721984) 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-MAPK6 / ERK3 antibody (HA721984) 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 HA721984, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of C2C12 cells labeling MAPK6 / ERK3 with Rabbit anti-MAPK6 / ERK3 antibody (HA721984) 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-MAPK6 / ERK3 antibody (HA721984) 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 HA721984, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of C6 cells labeling MAPK6 / ERK3 with Rabbit anti-MAPK6 / ERK3 antibody (HA721984) 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-MAPK6 / ERK3 antibody (HA721984) 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 HA721984, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 293T cells labeling MAPK6 / ERK3.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721984, 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).

This data was developed using HA721984, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of C6 cells labeling MAPK6 / ERK3.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721984, 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).