pEXP2-DEST Vector Kit

To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway™ destination vectors for expression in E. coli, insect, yeast,자세히 알아보기

카탈로그 번호	수량
V96002 V960-02으로도 사용됨	6 μg

카탈로그 번호 V96002

V960-02으로도 사용됨

제품 가격(KRW)

대량 주문 또는 맞춤형 요청

To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway™ destination vectors for expression in E. coli, insect, yeast, or mammalian cells, as well as for production of native protein or N- or C-terminal fusion proteins. All Gateway™ destination vectors have attR sites for recombination with any attL-flanked fragment, regardless of whether it is an entry clone or an Ultimate™ RF Clone. The following table lists the wide range of destination vectors available.

Additional materials required, available separately: Gateway™ entry clone, Gateway™ LR Clonase™ enzyme mix, and reaction buffer.

For Research Use Only. Not for use in diagnostic procedures.

항생제 내성 박테리아Ampicillin (AmpR), Zeocin™ (ZeoR)

제품 유형Gateway System Destination Expression Vector

수량6 μg

선택 제제(진핵)None

벡터pEXP, pDEST

클로닝 방법Gateway

제품라인Expressway, Gateway

프로모터T7

단백질 태그His Tag (6x), V5 Epitope Tag

Unit Size6 µg

구성 및 보관

All destination vectors are provided lyophilized and supercoiled.

자주 묻는 질문(FAQ)

I accidentally stored my E. coli slyD-Extract, E. coli Reaction Buffer (-A.A.), and 2X feed buffer at room temperature. Can I still use them?

Unfortunately, this may result in a loss of activity.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

I've run out of the T7 RNA polymerase for my cell-free expression. What do you suggest I use?

We would recommend using T7 RNA polymerase (Cat. No. 18033019, 50 U/µL). Use 1-1.5 µL in a 50 µL reaction system.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

I'm getting smearing after running my cell-free expression reaction on a gel.. What could be the cause of this?

Smearing may occur if samples for the following reasons:

- Samples were not precipitated with acetone: precipitate proteins with acetone to remove background smearing.
- Too much protein was loaded: reduce the amount used.
- The gel itself was not clean: rinse the gel briefly before exposing to film.
- Ethanol was present in the protein synthesis reaction: make sure that any residual ethanol is removed during DNA purification.
- Check the date of your pre-cast gels: do not use gels after the expiration date.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

I'm seeing a ladder of small-sized products after running my reaction on a gel when using the Expressway system. Why is this?

There may be several reasons for why this is occurring. The most common are: proteolysis, degradation of DNA and/or RNA templates (truncated templates will generate truncated protein products), internal initiation (if there are many methionines and internal RBS-like sequences in the gene, the ribosome may initiate translation from the wrong methionine), premature termination, translational pausing, frequent rare codon usage, complicated secondary structure of RNA, and others. This can also happen if proteins are denatured for too long, or not enough SDS was added to the 1X SDS-PAGE sample buffer.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

With a cell-free expression system, I'm getting good protein yield, but it has low biological activity. What can I do?

- Your protein may not be folding properly: try to reduce the incubation temperature to as low as 25 degrees C during synthesis.
- You may require post-translational modification of your protein: the Expressway system will not introduce post-translational modifications to the recombinant protein.
- Your synthetic protein may require co-factors for complete activity: try adding required co-factors to the protein synthesis reaction.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

View all pEXP2-DEST Vector Kit FAQs

인용 및 참조 문헌 (1)

인용 및 참조 문헌

Abstract

A Trim5-cyclophilin A fusion protein found in owl monkey kidney cells can restrict HIV-1.

Authors:Nisole S, Lynch C, Stoye JP, Yap MW,

Journal:Proc Natl Acad Sci U S A

PubMed ID:15326303

'Lv1 restriction of HIV-1 in the cells of Old World monkeys is associated with the expression of the Trim5 gene. Uniquely, in owl monkey kidney cells, HIV-1 restriction is dependent on the ability of incoming viral capsid protein to bind cyclophilin A (CypA). Cloning of the owl monkey Trim5 gene ... More