Rabbit MARK3 Polyclonal Antibody | anti-MARK3 antibody

Anti-MARK3 Antibody Picoband

Cat. Num. AAA19841

• Gene Names: MARK3; KP78; CTAK1; PAR1A; Par-1a
• Reactivity: Human, Mouse, Rat
• Applications: Western Blot, Immunohistochemistry, Flow Cytometry, Functional Assay, ELISA
• Purity: Immunogen affinity purified.
• Synonyms: MARK3; Polyclonal Antibody; Anti-MARK3 Antibody Picoband; Zinc finger protein Helios; aryl hydrocarbon receptor nuclear translocator like 2; Cytoskeleton-associated protein 5; Colonic and hepatic tumor overexpressed gene protein; Ch-TOG; CKAP5; KIAA0097; anti-MARK3 antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Expressed in fetal brain.Highly expressed in brain and placenta. Lower levels in heart, liver, thymus, kidney and lung. Located to endothelial cells and neuronal cells of the suprachiasmatic nucleus (SCN). Also detected in endothelial cells of the heart, lung and kidney. In the brain, specificallyExpressed in the thalamus, hippocampus and amygdala.
• Purity/Purification: Immunogen affinity purified.
• Form/Format: Lyophilized. Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

• Applicable Applications for anti-MARK3 antibody: Western Blot (WB), Immunohistochemistry (IHC), Flow Cytometry (FC/FACS), ELISA (EIA)
• Application Notes: WB: 0.25-0.5ug/ml, Human, Mouse, Rat; IHC(Paraffin-embedded Section): 2-5ug/ml, Human, Mouse, Rat; FC/FACS (Fixed): 1-3ug/1x10⁶ cells, Human; ELISA: 0.1-0.5ug/ml
• Immunogen: E coli-derived human MARK3 recombinant protein (Position: H311-D648).
• Subcellular Localization: Nucleus
• Reconstitution: Adding 0.2 ml of distilled water will yield a concentration of 500ug/ml.
• Cross Reactivity: No cross-reactivity with other proteins.
• Protein Function: Transcriptional activator which forms a core component of the circadian clock. The circadian clock, an internal time-keeping system, regulates various physiological processes through the generation of approximately 24 hour circadian rhythms in gene expression, which are translated into rhythms in metabolism and behavior. It is derived from the Latin roots 'circa' (about) and 'diem' (day) and acts as an important regulator of a wide array of physiological functions including metabolism, sleep, body temperature, blood pressure, endocrine, immune, cardiovascular, and renal function. Consists of two major components: the central clock, residing in the suprachiasmatic nucleus (SCN) of the brain, and the peripheral clocks that are present in nearly every tissue and organ system. Both the central and peripheral clocks can be reset by environmental cues, also known as Zeitgebers (German for 'timegivers'). The predominant Zeitgeber for the central clock is light, which is sensed by retina and signals ly to the SCN. The central clock entrains the peripheral clocks through neuronal and hormonal signals, body temperature and feeding-related cues, aligning all clocks with the external light/dark cycle. Circadian rhythms allow an organism to achieve temporal homeostasis with its environment at the molecular level by regulating gene expression to create a peak of protein expression once every 24 hours to control when a particular physiological process is most active with respect to the solar day. Transcription and translation of core clock components (CLOCK, NPAS2, ARNTL/BMAL1, ARNTL2/BMAL2, PER1, PER2, PER3, CRY1 and CRY2) plays a critical role in rhythm generation, whereas delays imposed by post-translational modifications (PTMs) are important for determining the period (tau) of the rhythms (tau refers to the period of a rhythm and is the length, in time, of one complete cycle). A diurnal rhythm is synchronized with the day/night cycle, while the ultradian and infradian rhythms have a period shorter and longer than 24 hours, respectively. Disruptions in the circadian rhythms contribute to the pathology of cardiovascular diseases, cancer, metabolic syndromes and aging. A transcription/translation feedback loop (TTFL) forms the core of the molecular circadian clock mechanism. Transcription factors, CLOCK or NPAS2 and ARNTL/BMAL1 or ARNTL2/BMAL2, form the positive limb of the feedback loop, act in the form of a heterodimer and activate the transcription of core clock genes and clock-controlled genes (involved in key metabolic processes), harboring E-box elements (5'-CACGTG-3') within their promoters. The core clock genes: PER1/2/3 and CRY1/2 which are transcriptional repressors form the negative limb of the feedback loop and interact with the CLOCK|NPAS2-ARNTL/BMAL1|ARNTL2/BMAL2 heterodimer inhibiting its activity and thereby negatively regulating their own expression. This heterodimer also activates nuclear receptors NR1D1/2 and RORA/B/G, which form a second feedback loop and which activate and repress ARNTL/BMAL1 transcription, respectively. The CLOCK-ARNTL2/BMAL2 heterodimer activates the transcription of SERPINE1/PAI1 and BHLHE40/DEC1.
• Preparation and Storage: At -20 degree C for one year from date of receipt. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for six months.Avoid repeated freezing and thawing.

FCM (Flow Cytometry)

(Figure 6. Flow Cytometry analysis of 293T cells using anti-MARK3 antibody (AAA19841).Overlay histogram showing 293T cells stained with AAA19841 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MARK3 Antibody (AAA19841, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IHC (Immunohistochemistry)

(Figure 5. IHC analysis of MARK3 using anti-MARK3 antibody (AAA19841).MARK3 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MARK3 Antibody (AAA19841) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MARK3 Antibody (AAA19841) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MARK3 Antibody (AAA19841) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MARK3 Antibody (AAA19841) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Lane 2: human K562 whole cell lysates,Lane 3: rat NRK whole cell lysates,Lane 4: mouse EL-4 whole cell lysates,Lane 5: mouse 4T1 whole cell lysates,Lane 6: mouse J774A.1 whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MARK3 antigen affinity purified polyclonal antibody (#AAA19841) at 0.5ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for MARK3 at approximately 80 kDa. The expected band size for MARK3 is at 80 kDa.)

Related Product Information for anti-MARK3 antibody

The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.

Product Categories/Family for anti-MARK3 antibody

Primary Antibodies; Cardiogenesis; Cardiovascular; Domain Families; Epigenetics and Nuclear Signaling; Hlh/Leucine Zipper; Neurology Process; Neuroscience; Receptors; Transcription; Transcription Factors; Transcription Factors/Regulators

References

1. Ansar, M., Chung, H., Waryah, Y. M., Makrythanasis, P., Falconnet, E., Rao, A. R., Guipponi, M., Narsani, A. K., Fingerhut, R., Santoni, F. A., Ranza, E., Waryah, A. M., Bellen, H. J., Antonarakis, S. E. Visual impairment and progressive phthisis bulbi caused by recessive pathogenic variant in MARK3. Hum. Molec. Genet. 27: 2703-2711, 2018.2. Lennerz, J. K., Hurov, J. B., White, L. S., Lewandowski, K. T., Prior, J. L., Planer, G. J., Gereau, R. W., IV, Piwnica-Worms, D., Schmidt, R. E., Piwnica-Worms, H. Loss of Par-1a/MARK3/C-TAK1 kinase leads to reduced adiposity, resistance to hepatic steatosis, and defective gluconeogenesis. Molec. Cell. Biol. 30: 5043-5056, 2010. 3. Muller, J., Ory, S., Copeland, T., Piwnica-Worms, H., Morrison, D. K. C-TAK1 regulates Ras signaling by phosphorylating the MAPK scaffold, KSR1. Molec. Cell 8: 983-993, 2001.

NCBI and Uniprot Product Information

• NCBI GI #: 193083125
• NCBI GeneID: 4140
• NCBI Accession #: NP_001122390.1
• NCBI GenBank Nucleotide #: NM_001128918.1
• UniProt Accession #: P27448; O60219; Q86TT8; Q8TB41; Q8WX83; Q96RG1; Q9UMY9; Q9UN34
• Molecular Weight: 84,489 Da
• NCBI Official Full Name: MAP/microtubule affinity-regulating kinase 3 isoform a
• NCBI Official Synonym Full Names: MAP/microtubule affinity-regulating kinase 3
• NCBI Official Symbol: MARK3
• NCBI Official Synonym Symbols: KP78; CTAK1; PAR1A; Par-1a
• NCBI Protein Information: MAP/microtubule affinity-regulating kinase 3; EMK-2; C-TAK1; ELKL motif kinase 2; protein kinase STK10; ser/Thr protein kinase PAR-1; cdc25C-associated protein kinase 1; serine/threonine-protein kinase p78
• UniProt Protein Name: MAP/microtubule affinity-regulating kinase 3
• UniProt Gene Name: MARK3
• UniProt Synonym Gene Names: CTAK1; EMK2; cTAK1; EMK-2; Par-1a
• UniProt Entry Name: MARK3_HUMAN