MARK3 Recombinant Rabbit Monoclonal Antibody [JE44-84]
cat.: ET7109-39
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IP, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: JE44-84
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. 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: 84 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human MARK3 aa 567-616 / 753.
Positive control: Rat brain tissue lysate, A431 cell lysate, 293 cell lysate, MCF-7, SH-SY5Y, SiHa, human appendix tissue, human breast cancer tissue, mouse small intestine tissue.
Subcellular location: Cell membrane. Peripheral membrane protein.
Recommended Dilutions:
  WB
  IP
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:500
1:10-1:50
1:50-1:200
1:50-1:200
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: P27448 Human | Q03141 Mouse | Q8VHF0 Rat
Alternative names: C-TAK1 Cdc25C associated protein kinase 1 Cdc25C-associated protein kinase 1 cTAK1 ELKL motif kinase 2 EMK-2 Emk2 ETK 1 KIAA4230 KP78 MAP microtubule affinity regulating kinase 3 MAP/microtubule affinity-regulating kinase 3 Mark3 MARK3_HUMAN Par 1a PAR1A Protein kinase STK10 Ser/Thr protein kinase PAR-1 Serine threonine protein kinase p78 Serine/threonine-protein kinase p78 SerThr protein kinase PAR 1
Images
ET7109-39_1.jpg Fig1: Western blot analysis of MARK3 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 was used at a 1:500 dilution 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: rat brain tissue lysate
Lane 2: A431 cell lysate
Lane 3: 293 cell lysate
ET7109-39_2.jpg Fig2: Immunocytochemistry staining MARK3 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 MARK3 monoclonal antibody at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
ET7109-39_3.jpg Fig3: Immunocytochemistry staining MARK3 in SH-SY5Y 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 MARK3 monoclonal antibody at a dilution of 1:50 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
ET7109-39_4.jpg Fig4: Immunocytochemistry staining MARK3 in SiHa 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 MARK3 monoclonal antibody at a dilution of 1:50 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
ET7109-39_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human appendix tissue using anti-MARK3 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 antibody (ET7109-39) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
ET7109-39_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue using anti-MARK3 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 antibody (ET7109-39) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
ET7109-39_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue using anti-MARK3 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 antibody (ET7109-39) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
ET7109-39_8.jpg Fig8: Flow cytometric analysis of MARK3 was done on MCF-7 cells. The cells were fixed, permeabilized and stained with MARK3 antibody at 1/100 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). After incubation of the primary antibody on room temperature for 1 hour, the cells was stained with a Alexa Fluor™ 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.