E. coli DNA Photolyase

Cat.No: FNK-01-013

Size: 20 μg

Concentration: 0.4 mg/ml 

Storage: -20℃. Avoid freeze-thaw cycles.

Purity: Over 95% by SDS-PAGE

Buffer: 25 mM Tris-HCI (pH 7.4), 50 mM KCl, 0.5 mM EDTA, 5 mM β-mercaptoethanol, 50% Glycerol Purity Over 95% by SDS-PAGE
 

Product Desrciption

Purified recombinant full-size E. coli Photolyase (Deoxyribodipyrimidine photo-lyase) as described in Sancar A, Smith FW, Sancar GB.

Purification of Escherichia coli DNA photolyase J Biol Chem. 1984 May 10;259(9):6028-32. PMID: 6325459 Length:472 aa. Mass (Da):53,667
 

Application

1. SDS-PAGE

2. Western blot

3. Immunoprecipitation

4. ELISA

5. Functional Studies
 

Background

Involved in repair of UV radiation-induced DNA damage. Catalyzes the light-dependent monomerization (300-600 nm) of cyclobutyl pyrimidine dimers (in cis-syn configuration), which are formed between adjacent bases on the same DNA strand upon exposure to ultraviolet radiation. Upon absorption of visible light electrons are transferred from Trp-307 through Trp-360 to Trp 383, and from there to FADH, giving rise to the fully reduced catalytic FADH-.
 

Data Link

UniProtKB: P00914 (PHR_ECOLI) 

Data Images

01-013 E. coli DNA Photolyase (Cyclobutane Pyrimidine Dimer Photolyase)

	Fig 1. SDS-PAGE analysis of E. coli Photolyase 1 μg of Photolyase was applied and analyzed on 12% SDS-PAEG.
	Fig.2 Western blot of E. coli photolyase Samples 1. Purified Photolyase 8.4 ng 2. Purified Photolyase 42 ng 3. E. coli crude lysate SDS-PAGE 12.5% gel Blotting: 15 v, overnight, Bloking by 5% skim milk Anti-Photolyase antibody used at 1/1,000 dilution. 2nd antibody: Goat anti-rabbit IgG (H & L) (ab97051) used at 1/10,000. Note that there are only 10-20 molecules of photolyase per E.coli cell.
	Fig 3 Immunoprecipitationm of E. coli Photolyase Immunoprecipitation was performed by using Dynabeads Magnetic system (Thermo Fisher). 1. E. coli AB1157 cell lysate (300 μg), antibody 2 μg 2. E. coli AB1157 cell lysate (300 μg), antibody 10 μg 3. Positve control: E. coli Photolyase purified, 4.2 μg, antibody 10 μg 4. Negative control: No sample, antibody 10 μg Western blot: Anti-Photolyase antibody used at 1/1,000 dilution and as the second antibody, HRP-conjugated goat anti-rabbit IgG VeriBlot for IP (ab131366) was used at 1/1,000 dilution.

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. NOT FOR MILITARY USE.