MAPK6/ERK3 Recombinant Rabbit Monoclonal Antibody [PSH03-39]
cat.: HA721984
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, FC
Clonality: Monoclonal
Clone number: PSH03-39
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 83 kDa
Isotype: IgG
Immunogen: Recombinant protein within human MAPK6 aa 1-350 / 721.
Positive control: HeLa cell lysate, A431 cell lysate, A549 cell lysate, PC-3M cell lysate, C2C12 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, 293T, C2C12, C6.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  FC

1:2,000
1:100
1:1,000
Uniprot #: SwissProt: Q16659 Human | Q61532 Mouse | P27704 Rat
Alternative names: ERK-3 ERK3 Extracellular signal regulated kinase 3 Extracellular signal regulated kinase p97 Extracellular signal-regulated kinase 3 HsT17250 MAP kinase 6 MAP kinase isoform p97 MAPK 6 MAPK6 Mitogen activated protein kinase 6 Mitogen-activated protein kinase 6 MK06_HUMAN p97 MAPK p97-MAPK PRKM6 Protein kinase mitogen activated 5 Protein kinase mitogen activated 6
Images
HA721984_1.jpg Fig1: Western blot analysis of MAPK6/ERK3 on different lysates with Rabbit anti-MAPK6/ERK3 antibody (HA721984) at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A431 cell lysate
Lane 3: A549 cell lysate
Lane 4: PC-3M cell lysate
Lane 5: C2C12 cell lysate
Lane 6: NIH/3T3 cell lysate
Lane 7: C6 cell lysate

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 5 minutes;

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 1/2,000 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.
HA721984_2.jpg Fig2: 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.
HA721984_3.jpg Fig3: 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.
HA721984_4.jpg Fig4: 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.
HA721984_5.jpg Fig5: 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).
HA721984_6.jpg Fig6: 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.