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Recombinant HIV1 Reverse Transcriptase protein (Active) (ab63979)

Submit a review Q&A (5)References (3)
SDS-PAGE - Recombinant HIV1 Reverse Transcriptase protein (ab63979)
  • Western blot - Recombinant HIV1 Reverse Transcriptase protein (ab63979)

Key features and details

  • Expression system: Escherichia coli
  • Purity: > 90% SDS-PAGE
  • Active: Yes
  • Suitable for: ELISA, SDS-PAGE, Functional Studies, WB

製品の詳細

  • 製品名

    Recombinant HIV1 Reverse Transcriptase protein (Active)

  • 生理活性

    5,000 units/ml.

    Definition of activity: Activity of 1 nanomole intake of dTMP in 10 min is considered 1 unit with poly(rA) and oligo(dT) as template and primer at 37°C.

  • 精製度

    > 90 % SDS-PAGE.
    (CBB staining) Fractionated by step-wise ammonium sulfate precipitation.

  • 発現系

    Escherichia coli

  • アクセッション番号

    P04585

  • タンパク質長

    Full length protein

  • Animal free

    No

  • 由来

    Recombinant

    • 生物種

      HIV-1

    • 配列の追加情報

      Composed of two subunits whose molecular weights are 66 kD and 51 kD, same as the those of the enzyme purified from AIDS virus particles. (GenBank AAA44988.1)

特性

Our Abpromise guarantee covers the use of ab63979 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • アプリケーション

    ELISA

    SDS-PAGE

    Functional Studies

    Western blot

  • 製品の状態

    Liquid

  • 備考

    ab63979 has been derived from infectious plasmid clone pNL4-3 and belongs to Group B of HIV-1.

    Applications:

    1) It is extremely effective for screening new specific inhibitors for HIV virus as a drug for treating AIDS.

    2) Generally, Gag and Env proteins are employed as antigens for detecting anti-HIV-1 antibody. However, by using this enzyme in combination as an antigen, the detection will be more sensitive.

    3) Reverse transcriptases are used in the first step of RT-PCR reaction for converting RNA to DNA. The HIV-1 reverse transcriptase can also be applied for RT-PCR method.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Concentration information loading...

前処理および保存

  • 保存方法および安定性

    Shipped at 4°C. Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.

    pH: 6
    Constituents: 0.1% Triton-X-100, 0.05% Magnesium chloride, 0.63% Tris HCl, 50% Glycerol, 0.29% Sodium chloride, 0.0154% DTT

    This product is an active protein and may elicit a biological response in vivo, handle with caution.

関連情報

  • 別名

    • HIV 1 Reverse transcriptase
    • HIV1 RT
    • Human Immunodeficiency virus 1
    • p51 RT
    • p66 RT
    • Reverse transcriptase p51 subunit
    • Reverse transcriptase p66 subunit
    see all

  • 関連性

    HIV1 Reverse Transcriptase is an RNA-dependent DNA polymerase of HIV1 (AIDS virus), subtype B origin. It is responsible for the production of a double stranded DNA copy of the single stranded RNA genome that is contained in the HIV1 virus particle and primed by the host cell lysine-tRNA which partly unfolds and anneals to the 5' end of the viral genomic RNA. This is extended by the polymerase function of RT to give a DNA-RNA hybrid. It also has RNaseH activity which degrades the RNA component of this hybrid once it has been copied.

画像

  • SDS-PAGE - Recombinant HIV1 Reverse Transcriptase protein (ab63979)
    SDS-PAGE - Recombinant HIV1 Reverse Transcriptase protein (ab63979)

    Polyacrylamide gel electrophoresis of ab63979.

  • Western blot - Recombinant HIV1 Reverse Transcriptase protein (ab63979)
    Western blot - Recombinant HIV1 Reverse Transcriptase protein (ab63979)
    All lanes : Anti-HIV1 Reverse Transcriptase antibody [39/4.12.2] (ab9066) at 1/2000 dilution

    Lane 1 : Recombinant HIV1 Reverse Transcriptase protein (Active) (ab63979) at 0.04 µg
    Lane 2 : Recombinant HIV1 Reverse Transcriptase protein (Active) (ab63979) at 0.02 µg
    Lane 3 : Recombinant HIV1 Reverse Transcriptase protein (Active) (ab63979) at 0.004 µg
    Lane 4 : Recombinant HIV1 Reverse Transcriptase protein (Active) (ab63979) at 0.002 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Developed using the ECL technique.

プロトコール

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

参考文献 (3)

ab63979 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab63979 は 3 報の論文で使用されています。

  • Holan M  et al. Synthesis and Anti-Hepatitis B Activities of 3'-Fluoro-2'-Substituted Apionucleosides. Molecules 27:N/A (2022). PubMed: 35458620
  • Liu S  et al. Tryptophan decreases the intensity of lipopolysaccharide-induced acute lung injury in a rat model. Amino Acids 52:1139-1147 (2020). PubMed: 32789611
  • Hughes AJ & Herr AE Microfluidic Western blotting. Proc Natl Acad Sci U S A : (2012). SDS-PAGE . PubMed: 23223527

レビューと Q&A

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レビューを送る 質問を送る

1-5 of 5 Abreviews or Q&A

Question

Are there 200 units per vial?

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Answer



Yes there are 200 units/vial.

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Question

Hello
How many μM and mg are in 200 units of HIV1 Reverse Transcriptase protein (Active) (ab63979)?


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Answer

Thank you for contacting us.
This can vary from lot to lot, but 1 vial of the current lot of HIV1 reverse transcriptase protein is 10 ug in 50 ul.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

I am interested in HIV1 Reverse Transcriptase protein (ab63979) if it functional in vitro? Can it synthesize DNA from RNA template? What kind of RNA it may be and should it include some specific motifs? What you recomend to use for RNA template?\

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Answer



ab63979 HIV1 Reverse Transcriptase protein has been functionally tested as noted on the online datasheet, which reads

"Specific activity determined as 10,000 to 20,000 units/mg. Definition of activity: Activity of 1 nanomole intake of dTMP in 10 min is considered 1 unit with poly(rA) and oligo(dT) as template and primer at 37°C."

Further experimental details include:
"Definition of activity: Activity of intake of 1 nmole of dTMP in 10 min at 37℃ is considered as 1 unit using poly(rA) and oligo(dT) as template and primer.

Conditions of measurement: 50 mM Tris-HCl (pH 8.3), 10 mM MgCl2, 50mM KCl, 3 mM DTT, 0.1% Nonidet P-40, 20 ug/ml poly(rA)・oligo(dT)12-18, , 0.5 mM dTTP (with [3H] dTTP,1x105 cpm) , and 10-50 units/ml reverse transcriptase."

Unfortunately, further details regarding use of this protein as a reverse transcriptase are not available.

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Question

Customer called to ask how the intake of dTMP was determined when determining the specific activity of this product.

From datasheet:

Specific activity determined as 10,000 to 20,000 units/mg. Definition of activity: Activity of 1 nanomole intake of dTMP in 10 min is considered 1 unit with poly(rA) and oligo(dT) as template and primer at 37°C.

Read More
Answer

Thank you for contacting Abcam regarding this product.
I have had the opportunity to speak with the lab regarding the specific activity of this product. Their response is as follows:
Definition of activity: Activity of intake of 1 nmole of dTMP in 10 min at 37℃ is considered as 1 unit using poly(rA) and oligo(dT) as template and primer.

Conditions of measurement: 50 mM Tris-HCl (pH 8.3), 10 mM MgCl2, 50mM KCl, 3 mM DTT, 0.1% Nonidet P-40, 20 ug/ml poly(rA)・oligo(dT)12-18, , 0.5 mM dTTP (with [3H] dTTP,1x105 cpm) , and 10-50 units/ml reverse transcriptase
These products are covered by our Abpromise guarantee. We provide scientific support, replacement or refund should this product not perform as indicated on the datasheet. More information on our Abpromise may be found at the following link:
https://www.abcam.com/index.html?pageconfig=abpromise
I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Question

How was the protein quantified?

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Answer

Thank you for contacting Abcam.

I have heard back from the lab and they have told me that the protein concentration was measured by BCA method using assay kit from Pierce and the concentration was also confirmed by OD280. They measured the concentration of the whole protein rather just the active domain.

Please let me know if there is anything else I can help you with.

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