E.coli LexA repressor functional

01-005 01-006

Backgrounds: LexA protein is the repressor of SOS regulon and binds specific DNA sequence called SOS box. It is used as a bait in yeast two-hybrid system.

Appications.
1. As an antigen of a control of bait protein expressionin western
blotting with anti-LexA antibody.
2. For the studies of SOS regulon.

Purity: Over 90% by SDS-PAGE (CBB staining)

Protein concentration: 0.8 mg/ml as measured by BCA method

Form: 50% glycerol 10 mM Tris-HCl (pH 7.5) 2 mM EDTA 100 mM NaCl 5 mM
mercaptoethanol

anti-Sua7p/TFⅡB (S. cerevisiae) rabbit serum

62-009

Storage temperature: Ship at 4℃and store at -20℃

Reactivity: S. cerevisiae Sua7 / TFIIB protein

Immunogen: Rcombinant His-tagged full-size Sua7 protein

Applications
1.Western blotting. (1/1000~1/5000)
2.Immunoprecipitation
3.Chromatin Immuno-Precipitation
4.ELISA

Form: 0.1% sodium azide added to the antiserum

Background: The fundamental transcription factor TFIIB has the characteristics of stabilizing the DNA binding of TATA box-binding protein (TBP) and binding directly to DNA by its conformational change. Also its N terminal region binds to the RNA channel of RNA polymerase undertaking a very important role in the determination of transcription initiation point and promoter clearance. Sua7p is the TFIIB of budding yeast and is composed of 346 amino acid residues (aa)

anti-Swi6 (S. pombe) antibody rabbit serum

Code:63-101

Product: Rabbit serum raised by immunizing full-size Swi6 protein of S. pombe expressed in E.coli

Application : For the studies of RNAi mechanism
1. Western blotting (x 2000~10000 dilution)
2. Immunoprecipitation

Specifications
State: Undiluted antiserum added with 0.09% sodium azide
Reactivity: Swi6 protein of S. pombe
References

1. Yao MC et al. Science 300:1581 (2003)
2.Ekwall K et al. Science 269:1429 (1995)
3. Wang G et al. Mol Cell Biol 20:6970 (2000)

Anti-histone H2B (S. pombe) antibody rabbit serum

Code:63-125 63-126

In the eukaryotic cells DNA is packaged repetitively into nucleosomes by means of interactions among two molecules of four classes of histone H2A H2B H3 and H4. Each of the histone proteins has an evolutionarily conserved amino-terminal ‘tail’ that protrudes from the nucleosome. This tail is the target of numerous diverse signaling pathways resulting in the addition of many post-translational modifications. These modifications include phosphorylation acetylation methylation ADP-ribosylation and mono-ubiquitination. Many important new modifications within the structured core and the carboxy-terminal tail regions of histones are also being identified. It is becoming increasingly clear that these modifications represent crucial regulatory events that govern the accessibility and function of the genome
Applications (see Ref 1)
1) Western blotting (1000 fold dilution). 2) Immunoprecipitation (CHIP assay).
Antigen: Synthetic peptide corresponding to the amino-terminal S. pombe histone H2B SAAEKKPASKAPAGKA
Reactivity: S. pombe histone H2B
Antibody: Undiluted rabbit antiserum added with 0.05 % sodium azide
Storage: -20℃ (long period -80℃)
Reference: This product has been used in the following reference.
1. Maruyama T. et al (Yanagida M). Histone H2B mutations in inner region affect ubiquitination centromere function silencing and chromosome segregation. EMBO J. 25: 2420-2431 (2006)

DNA (cytosine-5) methyltransferase (mouse) Dnmt1 active with reaction buffer and S-Adenosyl Methionine

code:10-201

DNA methylation is significant for epigenetic regulation of gene expression X chromosome inactivation genomic imprinting and development. Abberant methylation patterns are associated with certain human tumors and developmental abnormalities. In vertebrates there are two types of DNA methyltransferase activities; de novo and maintenance types. Two DNA methyltransferases Dnmt3a and Dnmt3b are responsible for the creation of methylation patterns at an early stage of embryogenesis (de novo-type) while Dnmt1 is responsible for the maintenance of methylation patterns during replication. Dnmt1 favors to methylate the hemimethylated DNA and preferentially methylates one strand of the double-stranded DNA during its processive methylation. This product mouse Dnmt1 deleting the N-terminal 290 amino acid residues was expressed using a baculovirus expression system and purified by Prof. S. Tajima and Dr. I. Suetake of Osaka University (ref.2). Since a trace of baculovirus contamination in the purified Dnmt1 the material and the container have to be sterilized at 121 C for 20 min before disposal according to Cartagena Protocol on Biodiversity.

Applications
1)In vitro metylation of cytosine residues in hemimethylated 　　　　　DNA at 5’….CG…3’. (ref. 12)
2)Antigen for anti-mammalian Dnmt1 antibodies.

Form: 0.5mg protein/ml in 0.2M NaCl 10mM HEPES (pH 7.4) 50% glycerol
Definition of specific activity: 1 unit is defined as the amount of the enzyme that transfer 1 pmole of
methyl group to poly dI-dC substrate during 30 minutes at 37℃

Specific activity：17 units/ul

Quality Assurance: Greater than 95% protein determined by SDS-PAGE (CBB staining) (Fig.1)

Reaction Conditions
Incubate in 1 x Dnmt1 Reaction Buffer (20mM Tris-HCl pH7.4 0.5 mM EDTA 0.2 mM DTT 5% glycerol) with 10μM S-adenosylmethionine (SAM) at 37℃