pBAD TOPO™ TA Expression Kit

본 제품은 LMO 제품으로, 고객 분께서 LMO 신고 시스템을 통해 직접 수입 신고를 진행해주셔야 합니다. 자세히보기

인용 및 참조 문헌 (2)

Invitrogen™

pBAD TOPO™ TA Expression Kit

Name: pBAD TOPO™ TA Expression Kit
SKU: K430040

pBAD TOPO™ TA Expression Kit는 Taq-amplified PCR 산물의 원스텝 클로닝과 원핵세포 발현을 위해 만들어졌습니다. TOPO™로 PCR 산물을 클로닝하면 바로 단백질을자세히 알아보기

보기 방식 변경

카탈로그 번호	단백질 태그	수량
K430040	V5 Epitope Tag	40 Reactions
K430001	His Tag (6x), V5 Epitope Tag	20 Reactions

2 개 옵션

카탈로그 번호 K430040

제품 가격(KRW)

단백질 태그:

V5 Epitope Tag

수량:

40 Reactions

pBAD TOPO™ TA Expression Kit는 Taq-amplified PCR 산물의 원스텝 클로닝과 원핵세포 발현을 위해 만들어졌습니다. TOPO™로 PCR 산물을 클로닝하면 바로 단백질을 발현할 수 있습니다. pBAD-TOPO™ vector의 편리한 특징:

• Taqamplified PCR 산물의 5분내 클로닝을 위한 linearized topoisomerase I-activated vector

Anti-V5 항체 검출을 위해 E. coli
V5 epitope tag에서 발현할 수 있도록 하는• araBAD promoter

• 니켈 킬레이트 수지를 사용한 정제 및 Anti-His(C-term) 항체 검출을 위한 C-terminal polyhistidine (6xHis) tag

• N-terminal leader peptide의 제거를 위한 Enterokinase 절단 부위

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

사양

항생제 내성 박테리아Ampicillin (AmpR)

박테리아 또는 효모 균주LMG194, TOP10

분열EK (Enterokinase) Recognition Site

구성 또는 유도성 시스템Inducible

발현 메커니즘Cell-Based Expression

발현 시스템E. coli

유도제Arabinose

제품 유형TA Expression Kit

수량40 Reactions

선택 제제(진핵)None

벡터pBAD

클로닝 방법TOPO-TA

제품라인TOPO

프로모터araBAD

단백질 태그V5 Epitope Tag

Unit SizeEach

구성 및 보관

Each pBAD TOPO™ TA Expression Kit contains two boxes and the LMG194 E. coli stab. The pBAD TOPO™ TA box contains all of the reagents necessary for PCR (except Taq polymerase), including 200 ng of topoisomerase I-activated pBAD-TOPO™ vector, sterile water, dNTPs, 10X PCR buffer, salt solution, control template and primers, 20% L-arabinose, pBAD primers for sequencing or PCR screening, and an expression control plasmid. Store at -20°C. The One Shot™ box contains all of the reagents necessary for transformation, including single-use 50 μl aliquots of One Shot™ TOP10 Chemically Competent E. coli, S.O.C. medium, and a supercoiled control plasmid. Store One Shot™ cells at -80°C. Store the LMG194 E. coli stab at at 2 - 8°C. All reagents are guaranteed stable for 6 months when properly stored.

자주 묻는 질문(FAQ)

Can I store my competent E. coli in liquid nitrogen?

We do not recommend storing competent E. coli strains in liquid nitrogen as the extreme temperature can be harmful to the cells. Also, the plastic storage vials are not intended to withstand the extreme temperature and may crack or break.

How should I store my competent E. coli?

We recommend storing our competent E. coli strains at -80°C. Storage at warmer temperatures, even for a brief period of time, will significantly decrease transformation efficiency.

How much L-arabinose should I use to induce expression with the pBAD expression system?

While the amount of L-arabinose can vary depending on your expression experiment, we suggest performing a pilot expression experiment with varying amounts of L-arabinose from 0.00002% to 0.2%.

Should I use TOP10 cells or the LMG194 E. coli strain you offer for expression with my pBAD system?

Top10

Advantages:
- Saves time, can go directly from cloning to expression.
- The glycerol stock is more stable because these strains are endA- and recA-.

Disdvantages:
- This strain is not protease-deficient. Therefore, the protein may be degraded.

LMG194

Advantages:
- Grows well in minimal media, except M9.
-Have to transform the plasmid into the cells just for expression.
-RM medium with glucose to ensure low basal level of protein.

Disadvantages:
- Not protease-deficient. Therefore, the protein may be degraded.
- The glycerol stock may not be stable because this cell strain is not recA- or endA-.

What competent cells do you recommend I use for expression with my pBAD expression system?

We recommend using a competent cell strain that is araBADC- and araEFGH+, allowing transportation of L-arabinose, but not metabolizing it. This is important for expression studies, as the level of L-arabinose will be constant inside the cell and will not decrease over time. We offer our TOP10 competent cells, or our LMG194 E. coli strain.

View all pBAD TOPO™ TA Expression Kit, 40 Reactions FAQs

인용 및 참조 문헌 (2)

인용 및 참조 문헌

Abstract

Overexpression, purification, and site-directed spin labeling of the Nramp metal transporter from Mycobacterium leprae.

Authors: Reeve Ian; Hummell David; Nelson Nathan; Voss John;

Journal:Proc Natl Acad Sci U S A

PubMed ID:12077319

'It has long been recognized that the pathogenicity of a broad range of intracellular parasites depends on the availability of transition metal ions, especially iron. Nramp1 (natural resistance-associated macrophage protein 1), a proton-coupled divalent metal ion transporter, has been identified as a controlling factor in the resistance or susceptibility to ... More

Ultrafast ligand rebinding in the heme domain of the oxygen sensors FixL and Dos: General regulatory implications for heme-based sensors.

Authors: Liebl Ursula; Bouzhir-Sima Latifa; Negrerie Michel; Martin Jean-Louis; Vos Marten H.;

Journal:Proc Natl Acad Sci U S A

PubMed ID:12271121

Heme-based oxygen sensors are part of ligand-specific two-component regulatory systems, which have both a relatively low oxygen affinity and a low oxygen-binding rate. To get insight into the dynamical aspects underlying these features and the ligand specificity of the signal transduction from the heme sensor domain, we used femtosecond spectroscopy ... More