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DNA Photolyase (E.coli)

AS21 4548 | Clonality: Polyclonal |  Host: Rabbit  | Reactivity: Escherichia coli

DNA Photolyase (E.coli) in the group Antibodies, Bacterial/Fungal at Agrisera AB (Antibodies for research) (AS21 4548)

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Product Information

Immunogen Purified, full length, recombinant DNA photolyase protein from E.coli, UniProt: P00914
Host Rabbit
Clonality Polyclonal
Purity Purified to a total IgG fraction using Protein A affinity.
Format Liquid in PBS with 50 % glycerol, filter sterlized
Quantity 200 µg
Storage Store at -20°C; make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications ELISA (ELISA), Immunoprecipitation (IP), Western blot (WB)
Recommended dilution assay dependent (ELISA), 1: 200 (IP), 1 : 1000 (WB)
Expected | apparent MW 53.6 kDa

Reactivity

Confirmed reactivity Escherichia coli
Predicted reactivity Species of your interest not listed? Contact us

Application examples

Application examples Western blot using anti-DNA photolyase (E.coli) antibodies

Samples: Purified E.coli photolyase 8.4 ng (1), Purified E.coli photolyase 42 ng (2), E. coli crude lysate, 40 μg (3) were separated on 12.5 % SDS-PAGE and transferred to a PVDF membrane at 15V, overnight, followed by blocking with 5 % non-fat milk, washes and incubation with a primary antibodies at 1: 1000 dilution 1h/RT in TBS-T, washes and incubation with secondary antibodies HRP conjugated. Reaction was visualized by chemiluminescnce following manufacture's recommendations.

Important note: 

there are only 10-20 molecules of photolyase per E.coli cell, therefore high protein load/well is necessary.

Immunoprecipitation using anti-DNA photolyase (E.coli) antibodies

Immunoprecipitation was performed by using Dynabeads Magnetic system (Thermo Fisher).
Samples: E. coli AB1157 cell lysate (300μg) , primary antibody 2μg (1),   E. coli AB1157 cell lysate (300μg), antibody 10 μg (2), Positve control: E. coli Photolyase purified, 4.2 μg, antibody 10 μg (3), Negative control: No sample, antibody 10 μg (4). Western blot: Anti-Photolyase antibody used at 1 : 1000 dilution and as the second antibody, HRP-conjugated goat anti-rabbit IgG for IP (suitable for detection of immunprecipitation products) was used at 1: 1000 dilution 1 h/RT. Reaction was visualized by chemiluminescence, following manufacture's recommendations.

Additional information

Related products

Background

Background Cyclobutane Pyrimidine Dimer photolyase (Deoxyribodipyrimidine photo-lyase) is 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 .

Product citations

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