Our broad portfolio consists of multiplex panels that allow you to choose, within the panel, analytes that best meet your needs. On a separate tab you can choose the premixed cytokine format or a single plex kit.
Cell Signaling Kits & MAPmates™
Choose fixed kits that allow you to explore entire pathways or processes. Or design your own kits by choosing single plex MAPmates™, following the provided guidelines.
The following MAPmates™ should not be plexed together:
-MAPmates™ that require a different assay buffer
-Phospho-specific and total MAPmate™ pairs, e.g. total GSK3β and GSK3β (Ser 9)
-PanTyr and site-specific MAPmates™, e.g. Phospho-EGF Receptor and phospho-STAT1 (Tyr701)
-More than 1 phospho-MAPmate™ for a single target (Akt, STAT3)
-GAPDH and β-Tubulin cannot be plexed with kits or MAPmates™ containing panTyr
.
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To begin designing your MILLIPLEX® MAP kit select a species, a panel type or kit of interest.
Custom Premix Selecting "Custom Premix" option means that all of the beads you have chosen will be premixed in manufacturing before the kit is sent to you.
If you have chosen panel analytes and then choose a premix or single plex kit, you will lose that customization.
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Add Additional Reagents (Buffer and Detection Kit is required for use with MAPmates)
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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
Space Saver Option Customers purchasing multiple kits may choose to save storage space by eliminating the kit packaging and receiving their multiplex assay components in plastic bags for more compact storage.
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RNA polymerase II (RNAPII or Pol II) is a multi-subunit enzyme responsible for the transcription of transcription of DNA into RNA from protein-coding genes. Transcription initiation requires recruitment of the complete transcription machinery to a promoter via solicitation by activators and chromatin remodeling factors. RNAPII can coordinate 10 to 14 subunits. This complex interacts with the promoter regions of genes and a variety of elements and transcription factors. The DNA binding domain of the polymerase is a groove where TFIIB orients the DNA for unwinding and transcription.
References
Product Information
Format
Purified
Control
Untreated and lambda phosphatase treated HeLa cell lysates.
Presentation
Purified rat monoclonal IgG2aκ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Detect the phospho-RNAPII (Thr4) protein using this Anti-phospho-RNAPII (Thr4), clone 1G7 validated for use in wb, IP, ICC & ChIP.
Key Applications
Western Blotting
Immunoprecipitation
Immunocytochemistry
Chromatin Immunoprecipitation (ChIP)
Application Notes
Immunoprecipitation Analysis: A representative lot from an independent laboratory detected phospho-RNAPII (Thr4) in HeLa cell extracts (Hintermair, C., et al. (2012). EMBO J. 31(12):2784-2791.).
Immunofluorescence Analysis: A representative lot from an independent laboratory detected phospho-RNAPII (Thr4) in HeLa cells (Hintermair, C., et al. (2012). EMBO J. 31(12):2784-2791.).
Chromatin Immunoprecipitation (ChIP) Analysis: A representative lot from an independent laboratory immunoprecipitated phospho-RNAPII (Thr4) in stably transfected Raji cell lysates (Hintermair, C., et al. (2012). EMBO J. 31(12):2784-2791.).
Biological Information
Immunogen
Ovalbumin-conjugated linear peptide corresponding to human RNAPII phosphorylated at Thr4.
Epitope
Phosphorylated Thr4
Clone
1G7
Host
Rat
Specificity
This antibody recognizes RNAPII phosphorylated at Thr4.
Evaluated by Western Blot in untreated and lambda phosphatase treated HeLa cell lysates.
Western Blot Analysis: A 1:500 dilution of this antibody detected phospho-RNAPII (Thr4) in 10 µg of untreated HeLa cell lysate. (Lambda phosphatase treatment was performed during the western blot procedure.)
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Threonine-4 of mammalian RNA polymerase II CTD is targeted by Polo-like kinase 3 and required for transcriptional elongation. Hintermair, Corinna, et al. EMBO J., 31: 2784-97 (2012)
2012
Eukaryotic RNA polymerase II (Pol II) has evolved an array of heptad repeats with the consensus sequence Tyr1-Ser2-Pro3-Thr4-Ser5-Pro6-Ser7 at the carboxy-terminal domain (CTD) of the large subunit (Rpb1). Differential phosphorylation of Ser2, Ser5, and Ser7 in the 5' and 3' regions of genes coordinates the binding of transcription and RNA processing factors to the initiating and elongating polymerase complexes. Here, we report phosphorylation of Thr4 by Polo-like kinase 3 in mammalian cells. ChIPseq analyses indicate an increase of Thr4-P levels in the 3' region of genes occurring subsequently to an increase of Ser2-P levels. A Thr4/Ala mutant of Pol II displays a lethal phenotype. This mutant reveals a global defect in RNA elongation, while initiation is largely unaffected. Since Thr4 replacement mutants are viable in yeast we conclude that this amino acid has evolved an essential function(s) in the CTD of Pol II for gene transcription in mammalian cells.
