ERK5 Recombinant Rabbit Monoclonal Antibody [SD2084]
cat.: ET1612-7
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IP, FC
Clonality: Monoclonal
Clone number: SD2084
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 88 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human ERK5 aa 767-816 / 816.
Positive control: HeLa cell lysate, NIH/3T3 cell lysate, Neuro-2a cell lysate, mouse brain tissue lysate, mouse testis tissue lysate, PC-12 cell lysate, NIH/3T3, PC-12, HeLa.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  FC
  IP
1:500-1:2,000
1:100
1:100-1:500
1:1,000
Use at an assay dependent concentration.
Uniprot #: SwissProt: Q13164 Human | Q9WVS8 Mouse | P0C865 Rat
Alternative names: Big MAP kinase 1 BMK 1 BMK 1 kinase BMK-1 BMK1 BMK1 Kinase EC 2.7.11.24 ERK 4 ERK 5 ERK-5 ERK4 ERK5 Extracellular signal regulated kinase 5 Extracellular signal-regulated kinase 5 MAP kinase 7 MAPK 7 MAPK7 Mitogen activated protein kinase 7 Mitogen-activated protein kinase 7 MK07_HUMAN OTTHUMP00000065906 OTTHUMP00000065907 PRKM 7 PRKM7 PROTEIN KINASE, MITOGEN-ACTIVATED, 7
Images
ET1612-7_1.jpg Fig1: Western blot analysis of ERK5 on different lysates with Rabbit anti-ERK5 antibody (ET1612-7) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: NIH/3T3 cell lysate (20 µg/Lane)
Lane 3: Neuro-2a cell lysate (20 µg/Lane)
Lane 4: Mouse brain tissue lysate (40 µg/Lane)
Lane 5: Mouse testis tissue lysate (40 µg/Lane)
Lane 6: PC-12 cell lysate (20 µg/Lane)

Predicted band size: 88 kDa
Observed band size: 115 kDa

Exposure time: 3 minutes; 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 (ET1612-7) at 1/1,000 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.
ET1612-7_2.jpg Fig2: Western blot analysis of ERK5 on different lysates with Rabbit anti-ERK5 antibody (ET1612-7) at 1/2,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si ERK5 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 88 kDa
Observed band size: 115 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 (ET1612-7) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1612-7_3.jpg Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling ERK5 with Rabbit anti-ERK5 antibody (ET1612-7) 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-ERK5 antibody (ET1612-7) 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.
ET1612-7_4.jpg Fig4: Immunocytochemistry analysis of PC-12 cells labeling ERK5 with Rabbit anti-ERK5 antibody (ET1612-7) 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-ERK5 antibody (ET1612-7) 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.
ET1612-7_5.jpg Fig5: Flow cytometric analysis of HeLa cells labeling ERK5.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1612-7, 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).
ET1612-7_6.jpg Fig6: Flow cytometric analysis of NIH/3T3 cells labeling ERK5.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1612-7, 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.