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-E.coli LexA antibody 　rabbit serum

Code:61-001

E. coli LexA protein binds specifically to the SOS-box sequence and represses the genes belonging to the SOS regulon. In response to DNA damage RecA protein is activated by ss-DNA accumulated in the damaged cells and promotes autocleavage of LexA repressor by its coprotease activity. As a result DNA repair genes and error prone polymerases are induced and DNA damage is repaired and mutation is induced (1).
The lexA gene is used for yeast two-hybrid experiments as a bait to identify the protein-protein interaction in vivo (2).
This product was prepared by immunizing rabbit with full-size highly-purified recombinant LexA protein. Using this antibody 23 kD LexA protein was identified in the E. coli whole-cell lysate (Fig 1) and the expression of bait constructs was identified in yeast extracts by Western blotting.

Applications
1) Studies on the SOS regulation in E .coli (3). For Western blotting; 1000~3000 fold dilution.
2) Construction and expression of a bait protein fused to LexA protein can be examined by Western blotting of the yeast extractsusing the antiserum.
Purified LexA protein is available from BioAcademia（#01-002）to be used as a positive control for Western blotting.
3) Immunohistochemistry (LexA fusion protein was detected in
transgenic Drosophila after fixation with 4% formalodehyde.)
4) Immunoprecipitation and chromatin immuno-precipitation

E.coli RuvA Protein functional

01-007 01-008

Product: Recombinant RuvA protein with no tag attached
Function:Holliday junction-specific binding protein. Forms a complex
with RuvB motor protein and together promotes branch
migration of Holliday junction in late stage of homologous
recombination.
Purity: RuvA protein over 90% by SDS-PAGE (CBB staining)
Concentration: 2.7 mg/ml (determined by BCA method)
Form: 50% glycerol 10 mM Tris-HCl (pH7.5) 2 mM EDTA 100 mM NaCl
5 mM mercaptoethanol
Storage: -20C (long period -80C)

DNA polymerase β (Rat) functional

Code:10-101

Backgrounds:DNA polymeraseβ is a distributive polymerase involved in base excision repair which repairs damaged DNA by excising modified bases (oxidized methylated deaminated etc.) (ref. 1).This product is highly purified full-length rat DNA polymeraseβ overproduced in E. coli with high enzymatic activity without any tag attached (ref.2).

Applications
1) For the studies on the mechanisms of base-excision repair of DNA damage
2) As a positive control of western blotting with anti-DNA polymeraseβ antibody

Properties of the product
Enzyme activity: 90 unit/ul (1unit of the enzyme activity incorporates 1 nanomole of dNTP into acid-insoluble fraction at 37℃ in 60 min.)
Purity: Over 95% pure by SDS-PAGE analysis
Form: 1.3 mg/ml in 50mM Tris-HCl pH7.6 0.3M KCl 0.1mM EDTA 1mM DTT 20% glycerol

References: This product is described in ref. 2
1. Friedberg EC et al. DNA Repair and Mutagenesis 2nd ed. ASM Press (2006)
2. Date T. et al. Biochemistry 27: 2983 (1988)

Related product
#70-041 anti-DNA polymeraseβ (rat) antibody affinity purified cross-reacts with human and mouse homologs.

anti-XPA(human)antibody monoclonal

code:70-031

XP (Xeroderma pigmentosum) is an autosomal recessive human disease characterized by hypersensitivity to sunlight and a high incidence of skin cancer on sun-exposed skin (1). Cells from XP patients are hypersensitive to killing by UV irradiation because of a defect in nucleotide excision repair (NER). XP is classified into seven complementation groups (A~G) and a variant form (1). XPA shows the most severe symptoms. Products encoded by the XP genes function in repairing UV-induced cyclobutane pyrimidine dimmer and (6-4) photoproducts as well as chemically induced variety of DNA lesions (1).
XPA protein consists of 273 amino acids and forms a complex with many proteins such as RPA ERCC1 TFIIH、XAB1 and XAB2 which plays a role in early step of NER. The hybridoma 5F12 was constructed by Prof. K. Tanaka’s group who first cloned the XPA gene (2 3).

Anti-DNA polymerase β antibody (rabbit antiserum)

70-041

Reactivity: human rat mouse hamster

Immunogen: Recombinant rat DNA polymerase beta functional

Applications
1) Western blotting (1/ 1000~ 1/3000)
2) Immunoprecipitation (1/100)
3) Immunofluorescent staining (1/1000)
4) ELISA (assay dependent)

Form: 1 mg/ml in PBS 50% glycerol filter-sterilized. Azide and carrier free

Anti-Cdc22 / RMM1 (S. pombe) antibody rabbit polyclonal affinity-pure

63-011

　The cdc22 gene of S. pombe encodes ribonucleoside-diphosphate reductase large chain (811 aa92 kDa) which forms heterodimer with the small chain. The heterodimer catalyzes the　biosynthesis of　deoxyribonucleotides from the corresponding ribonucleotides and provides theprecursors necessary for DNA synthesis.

Applications: Western blottind (1/1000 dilution.

Immunogen: Synthetic peptide NVNPTDLWDWAE-C corresponding to
571-582 residues of S. pombe Cdc22 protein.

Purity: Affinity purified with the immunogen peptide conjugated with agarose beads.

Form: 1 mg/ml in PBS with 50% glycerol. Filter-sterilyzed.

Storage: Sent at 4℃or -20℃. Upon arrival spin-down and store at -20℃.