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MARK3 Recombinant Rabbit Monoclonal Antibody [JE44-84]

Usd: 385

Specification

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Catalog# ET7109-39

MARK3 Recombinant Rabbit Monoclonal Antibody [JE44-84]

Application

  • WB

  • IP

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

MARK3 Recombinant Rabbit Monoclonal Antibody [JE44-84]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human MARK3 aa 567-616 / 753.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IP, IF-Cell, IF-Tissue, IHC-P, FC

Molecular Weight

Predicted band size: 84 kDa

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.

Conjugation

unconjugated

Clone Number

JE44-84

RRID

AB_3070926

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500

  • IP

  • 1:10-1:50

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

Target

Function

MARK3 (MAP/microtubule affinity-regulating kinase 3), also known as C-TAK1 (Cdc25C-associated protein kinase 1), EMK-2 (ELKL motif kinase 2), protein kinase STK10 or serine/threonine-protein kinase p78, is a 753 amino acid peripheral membrane protein that phosphorylates Cdc25C on Serine 216 and is ubiquitously expressed in various human tissue and cell lines. Existing as six alternatively spliced isoforms, MARK3 belongs to the Ser/Thr protein kinase family and the protein kinase superfamily. MARK3 has been suggested to mediate the binding of the 14-3-3 proteins through its kinase activity and acts as a negative regulator of mitosis. The gene encoding MARK3 maps to human chromosome 14q32.3 and mouse chromosome 12 F1.

Background References

1. Schwarz T et al. C-TAK1 interacts with microphthalmia-associated transcription factor, Mitf, but not the related family member Tfe3. Biochem Biophys Res Commun 394(4):890-5 (2010).

2. Al-Hakim AK et al. Control of AMPK-related kinases by USP9X and atypical Lys(29)/Lys(33)-linked polyubiquitin chains. Biochem J 411(2):249-60 (2008).

Sequence Similarity

Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. SNF1 subfamily.

Tissue Specificity

Ubiquitous.

Post-translational Modification

Phosphorylated at Thr-211 by STK11/LKB1 in complex with STE20-related adapter-alpha (STRADA) pseudo kinase and CAB39. Phosphorylation at Thr-564 by PRKCZ/aPKC inhibits the kinase activity.

Subcellular Location

Cell membrane. Peripheral membrane protein.

UNIPROT

SwissProt: P27448 Human

SwissProt: Q03141 Mouse

SwissProt: Q8VHF0 Rat

Synonyms

C-TAK1 antibody

Cdc25C associated protein kinase 1 antibody

Cdc25C-associated protein kinase 1 antibody

cTAK1 antibody

ELKL motif kinase 2 antibody

EMK-2 antibody

Emk2 antibody

ETK 1 antibody

KIAA4230 antibody

KP78 antibody

Expand

Images

  • 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 (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:5,000 dilution was used for 1 hour at room temperature.<br />Positive control: <br />Lane 1: rat brain tissue lysate<br />Lane 2: A431 cell lysate<br />Lane 3: 293 cell lysate<br />

    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

  • 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&trade; 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

    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).

  • 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&trade; 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

    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).

  • 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&trade; 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

    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).

  • 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 ddH<sub>2</sub>O and PBS, and then probed with the antibody (<a href="/products/ET7109-39" style="font-weight: bold;text-decoration: underline;">ET7109-39</a>) 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.

    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.

  • 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 ddH<sub>2</sub>O and PBS, and then probed with the antibody (<a href="/products/ET7109-39" style="font-weight: bold;text-decoration: underline;">ET7109-39</a>) 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.

    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.

  • 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 ddH<sub>2</sub>O and PBS, and then probed with the antibody (<a href="/products/ET7109-39" style="font-weight: bold;text-decoration: underline;">ET7109-39</a>) 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.

    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.

  • 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&trade; 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.

    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.

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