MEKK3 Recombinant Rabbit Monoclonal Antibody [SD0839]
cat.: ET1612-98
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: SD0839
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: 71 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human MEKK3 aa 1-50 / 626.
Positive control: 293T cell lysate, A431 cell lysate, Hela, MCF-7, SKOV-3, mouse uterus tissue, rat uterus tissue.
Subcellular location: Cytosol, cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:500-1:2,000
1:100-1:500
1:100-1:500
1:50-1:200
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: Q99759 Human | Q61084 Mouse
Unigene: 72680 Rat
Alternative names: M3K3_HUMAN MAP/ERK kinase kinase 3 MAP3K3 MAPK/ERK kinase kinase 3 MAPKKK3 MEK kinase 3 MEKK 3 MEKK3 Mitogen activated protein kinase kinase kinase 3 Mitogen-activated protein kinase kinase kinase 3
Images
ET1612-98_1.jpg Fig1: Western blot analysis of MEKK3 on different lysates with Rabbit anti-MEKK3 antibody (ET1612-98) at 1/2,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si MEKK3 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 71 kDa
Observed band size: 71 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 (ET1612-98) 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.
ET1612-98_2.jpg Fig2: Western blot analysis of MEKK3 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1612-98, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: 293T cell lysate
Lane 2: A431 cell lysate
ET1612-98_3.jpg Fig3: ICC staining of MEKK3 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-98, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1612-98_4.jpg Fig4: ICC staining of MEKK3 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-98, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1612-98_5.jpg Fig5: ICC staining of MEKK3 in SKOV-3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-98, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1612-98_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse uterus tissue using anti-MEKK3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-98, 1/50) for 30 minutes 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.
ET1612-98_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat uterus tissue using anti-MEKK3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-98, 1/50) for 30 minutes 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.
ET1612-98_8.jpg Fig8: Flow cytometric analysis of MEKK3 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1612-98, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.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”.