The new website is online and in the testing stage. We apologize for the inconvenience caused. Welcome to send the feedbacks to Levi@hongene.com. Thank you very much~

EN CN JP
account_circle login
home Home navigate_next Product navigate_next Enzyme navigate_nextFor RNA Synthesisnavigate_nextRNA Synthesis

Alkaline Phosphatase,TAB5

  • Cat.No.ON-179
  • SourceRecombinant E.coli
  • Purity>90% by SDS-PAGE
  • Storage-20℃
add_shopping_cart Add to Cart commentConsulting

Product description: Alkaline Phosphatase, TAB5 nonspecifically catalyzes the dephosphorylation of 5' and 3' ends of DNA and RNA phosphomonoesters. It also hydrolyses NTPs and dNTPs. It is completely inactivated by heating at 80℃ for 2 minutes.

Source: Recombinant E.coli

Concentration and Size: 2.5U/μL

Unit Definition: One unit is defined as the amount of enzyme that will dephosphorylate 1μg of pUC19 vector DNA cut with a restriction enzyme generating 5' recessed ends in 30 minutes at 37℃. Dephosphorylation is defined as > 95% inhibition of recircularization in a self-ligation reaction and is measured by transformation into DH5α.

Storage Buffer: 10mM Tris-HCl , 1mM MgCl2, 0.01mM ZnCl2, 50%(v/v) glycerol, pH7.4, 25℃

10X Phosphatase Reaction Buffer: 500mM Bis-Tris-Propane-HCl , 10mM MgCl2, 1mM ZnCl2, pH6, 25℃

10X IVT Reaction Buffer without Mg2+ , Cat#ON-41, 400mM Tris PH7.9, 100mM DTT, 20mM Spermidine


Related Products:

High Yield T7 RNA Synthesis Kit, Cat#ON-040

T7 RNA polymerase, Cat#ON-004

T4 RNA ligase 1 (ssRNA ligase), Cat#ON-081

Ribonuclease Inhibitor, Human Placenta, Cat#ON-039

10X Phosphatase Reaction Buffer, Cat#ON-180

Protocol:

DNA as substance:

1. Combine the follows on ice,

5'-Phosphate-DNA1μg
10X Phosphatase Reaction Buffer2μL
Alkaline Phosphatase, TAB52.5~5 units
Nuclease-free H2OTo 20μL

2. Incubate at 37℃, 30min

3. (optional) Heat inactivity enzyme by incubating at 80℃, 2min.

RNA as substance:

1. Heat Denature the RNA,

5'-Phosphate-RNA1μg
Nuclease-free H2OTo 15μL

2. Incubate at 65℃, 10~15min,  transfer the tube immediately to ice.

3. On ice, combine the following reaction components in the order given

15μLHeat denatured RNA from step 1 (above)
2μL10X Phosphatase Reaction Buffer
2.5~5 unitsAlkaline Phosphatasae, TAB5
20 unitsRNase inhibitor (Recommend)
20μLTotal reaction volume


4. Incubate at 37℃, 30min


5. Stop the reaction using any one of the following methods:

a) immediate storage at –20℃ or –70℃.

b) Add of EDTA to a final concentration of > 2 mM.

c) Phenol/chloroform extract then precipitate using salt/alcohol.

Storage Conditions: -20℃

Quality Assurance: Free of endonuclease, exonuclease, nickase and RNase activities.

Physical Purity: >90% by SDS-PAGE.

1. Maria Rina, Charalambos Pozidis, et al. Eur. J. Biochem. 267, 1230-1238 (2000).

2. Ellen Wang, Dimitris Koutsioulis, et al. J. Mol. Biol. 366, 1318-1331(2007).

3. Ping Li, Jon Beckwith, et al. Proc. Natl. Acad. Sci. USA.85,7685-7689(1988).

keyboard_arrow_up
US headquarters:

+1 510 931 4711

Sales Inquiry Ext. 303 / Shipping Logistics Ext. 304 / General Ext. 305 mailbox
Primary contact: product.info@hongene.com
Secondary contact: sales@hongeneusa.com
China Mainland Contact: sales@hongene.com
Address 29520 Kohoutek Way, union city CA.94587
copyright © 2019 Hongene All Rights Reserved

Login

close

register

close
shopping_cart

Shopping Cart

check_circleSubmit Order