pSecTag2 A, B, & C Mammalian Expression Vectors
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Citas y referencias (20)
Invitrogen™

pSecTag2 A, B, & C Mammalian Expression Vectors

pSecTag2 y pSecTag2/Hygro son vectores de expresión de mamíferos diseñados para la secreción, purificación y detección de proteínas de fusión.Más información
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Número de catálogoCantidad
V9002020μg
Número de catálogo V90020
Precio (CLP)
1.452.434
20 µg
Añadir al carro de la compra
Cantidad:
20μg
Precio (CLP)
1.452.434
20 µg
Añadir al carro de la compra
pSecTag2 y pSecTag2/Hygro son vectores de expresión de mamíferos diseñados para la secreción, purificación y detección de proteínas de fusión. Cada vector tiene un sitio grande de clonación múltiple en tres marcos de lectura para simplificar la clonación en marco con señal de secreción N-terminal. Los vectores (figura 1) ofrecen las siguientes funciones:

• Señal de secreción de la región V-J2-C de la cadena kappa de la Ig de ratón para una secreción eficaz de proteínas recombinantes (figura 2)
• Promotor de citomegalovirus (CMV) para una expresión constitutiva de alto nivel
• Etiqueta polihistidina (6xHis) C-terminal para una rápida purificación con resina quelante de níquel y detección con anticuerpos anti-His (C-terminal)
• Epítopo C-terminal c-myc para la detección con un anticuerpo Anti-myc
• Secuencia de terminación de la transcripción y señal de poliadenilación de la hormona de crecimiento bovino (BGH) para mejorar la estabilidad del ARNm
• Origen de replicación episomal SV40 y rescate de vector sencillo en líneas celulares que expresan el antígeno T grande (es decir, COS-1, COS-7)

Los vectores pSecTag2 portan el gen de resistencia a Zeocin™ para permitir una selección rentable en células de mamífero. La selección con Zeocin™ también puede usarse en E. coli.

Los vectores pSecTag2/Hygro tienen un gen de resistencia a la higromicina B para la selección de líneas celulares de mamífero estables.
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
Tipo de entregaTransfección
Para utilizar con (aplicación)Expresión secretada
Tipo de productoVector de expresión de mamíferos
Cantidad20μg
Agente de selección (eucariótico)Zeocin™
VectorpSec
Método de clonaciónEnzimas de restricción/MCS
PromotorCMV
Etiqueta de proteínaEtiqueta His (6x), Etiqueta de epítopo c-Myc, Secuencia líder de IgK
Unit Size20 µg
Contenido y almacenamiento
Se proporcionan 20 μg de cada vector y un control de expresión PSA en forma de ADN superenrollado y liofilizado. Conservar a -20°C. Se garantiza la estabilidad de los vectores durante 6 meses si se almacenan correctamente.

Preguntas frecuentes

I used the pSecTag2 vector and am seeing cellular but not secreted expression of the protein of interest. Can you offer some tips?

If the gene of interest has a start codon in the context of a perfect Kozak sequence, it may be preferentially translated over the vector’'s initiation codon, resulting in no leader sequence and no glycosylation, and hence no targeting to the endoplasmic reticulum and no secretion. This is rare, but it has been observed. If it is a problem, we recommend using PCR to delete the start codon.

Can I propagate my pSecTag2 vector in E. coli containing the Tn5 gene?

pSectag2 vectors have the Zeocin antibiotic-resistance marker for selection in E. coli, and any E. coli strain that contains the complete Tn5 transposable element (i.e., DH5alphaF'IQ, SURE, SURE2) encodes the ble (bleomycin) resistance gene that confers resistance to the Zeocin antibiotic. Hence, for the most efficient selection, we highly recommend choosing an E. coli strain that does not contain the Tn5 gene.

I see that the pSectag2 vectors carry the Zeocin antibiotic-resistance gene driven by the EM7 promoter. Can I use Zeocin antibiotic for selection when I propagate these vectors in E. coli?

Yes, you can use Zeocin antibiotic for selection in E. coli. However, keep in mind that for Zeocin antibiotic to be active, the salt concentration of the medium must remain low (<90 mM) and the pH must be 7.5. Prepare LB broth and LB agar plates using low-salt (5 g NaCl/liter) LB.

What is the difference between pSecTag2 and pSecTag2/Hygro vectors?

The only difference between these vectors is that the pSecTag2 vectors have the Zeocin antibiotic-resistance gene for stable selection, whereas the pSecTag2/Hygro vectors have the Hygromycin B resistance gene for stable selection.

I would like to get secreted expression of my protein. What are your recommendations?

You can insert an N-terminal secretion signal or leader sequence upstream of your gene and in-frame with the gene sequence to facilitate secreted expression of the protein. We actually offer the pSecTag2 (Cat. No. V90020) and pSecTag2/Hygro (Cat. No. V91020) vectors designed for this purpose. These vectors contain the N-terminal murine Ig kappa-chain secretion signal for secreted expression of the protein of interest.

View all pSecTag2 A, B, & C Mammalian Expression Vectors FAQs

Citations & References (20)

Citations & References
Abstract
Neuropilin-1 binds vascular endothelial growth factor 165, placenta growth factor-2, and heparin via its b1b2 domain.
Authors: Mamluk Roni; Gechtman Ze'ev; Kutcher Matthew E; Gasiunas Nijole; Gallagher John; Klagsbrun Michael;
Journal:J Biol Chem
PubMed ID:11986311
'Neuroplin-1 (NRP1), a receptor for vascular endothelial growth factor (VEGF) family members, has three distinct extracellular domains, a1a2, b1b2, and c. To determine the VEGF(165) and placenta growth factor 2 (PlGF-2)-binding sites of NRP1, recombinant NRP1 domains were expressed in mammalian cells as Myc-tagged, soluble proteins, and used in co-precipitation ... More
Molecular dissection of the alpha-dystroglycan- and integrin-binding sites within the globular domain of human laminin-10.
Authors:Ido H, Harada K, Futaki S, Hayashi Y, Nishiuchi R, Natsuka Y, Li S, Wada Y, Combs AC, Ervasti JM, Sekiguchi K,
Journal:J Biol Chem
PubMed ID:14701821
'The adhesive interactions of cells with laminins are mediated by integrins and non-integrin-type receptors such as alpha-dystroglycan and syndecans. Laminins bind to these receptors at the C-terminal globular domain of their alpha chains, but the regions recognized by these receptors have not been mapped precisely. In this study, we sought ... More
Sulfation of N-acetylglucosamine by chondroitin 6-sulfotransferase 2 (GST-5).
Authors: Bhakta S; Bartes A; Bowman K G; Kao W M; Polsky I; Lee J K; Cook B N; Bruehl R E; Rosen S D; Bertozzi C R; Hemmerich S;
Journal:J Biol Chem
PubMed ID:10956661
'Based on sequence homology with a previously cloned human GlcNAc 6-O-sulfotransferase, we have identified an open reading frame (ORF) encoding a novel member of the Gal/GalNAc/GlcNAc 6-O-sulfotransferase (GST) family termed GST-5 on the human X chromosome (band Xp11). GST-5 has recently been characterized as a novel GalNAc 6-O-sulfotransferase termed chondroitin ... More
Plexin-B1 directly interacts with PDZ-RhoGEF/LARG to regulate RhoA and growth cone morphology.
Authors: Swiercz Jakub M; Kuner Rohini; Behrens Jürgen; Offermanns Stefan;
Journal:Neuron
PubMed ID:12123608
'Plexins are widely expressed transmembrane proteins that, in the nervous system, mediate repulsive signals of semaphorins. However, the molecular nature of plexin-mediated signal transduction remains poorly understood. Here, we demonstrate that plexin-B family members associate through their C termini with the Rho guanine nucleotide exchange factors PDZ-RhoGEF and LARG. Activation ... More
Death receptor 5, a new member of the TNFR family, and DR4 induce FADD- dependent apoptosis and activate the NF-kappaB pathway.
Authors:Chaudhary PM, Eby M, Jasmin A, Bookwalter A, Murray J, Hood L
Journal:Immunity
PubMed ID:9430227
'Death receptor 4 (DR4) is a recently described receptor for the cytotoxic ligand TRAIL that reportedly uses a FADD-independent pathway to induce apoptosis and does not activate the NF-kappaB pathway. We have isolated a new member of the tumor necrosis factor receptor (TNFR) family, designated DR5, which bears a high ... More
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