Building of a GFP/Puro double-labeled lentiviral vector containing CK8 interfering RNA and its impact on cell apoptosis in vitro
OBJECTIVE
To assemble a GFP/Puro double-labeled lentiviral expression vector for CK8 silencing and assess the consequences of CK8 silencing on cell apoptosis.
METHODS
The siRNA sequences of CK8 have been inserted into the lentiviral expression vector GV248 and transfected into 293T cells with the packaging plasmids PMD and SPA. The lentivirus was collected at 24 and 36 h post-transfection. Circulate cytometry was used to detect the virus titer and the optimistic cells have been chosen with puromycin. The knockdown of CK8 was examined by Western blotting. The impact of CK8 down-regulation on cell apoptosis induced by cisplatin was detected with Annexin V/PI staining.
CONCLUSIONS
We efficiently constructed CK8 interference lentiviral vector and obtained a steady cell line with CK8 knock-down that was delicate to cisplatin-induced apoptosis
gfpvector
 anti-SPOCK1 |
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| YF-PA14763 | Abfrontier | 50 ug | EUR 363 |
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Description: Mouse polyclonal to SPOCK1 |
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 SPOCK1 ELISA KIT|Human |
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| EF003197 | Lifescience Market | 96 Tests | EUR 689 |
 Human SPOCK1 shRNA Plasmid |
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| 20-abx954565 | Abbexa |
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) SPOCK1 Recombinant Protein (Human) |
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| RP029953 | ABM | 100 ug | Ask for price |
 (pORF)) Spock1 ORF Vector (Mouse) (pORF) |
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| ORF058387 | ABM | 1.0 ug DNA | EUR 506 |
Spock1 ORF Vector (Mouse) (pORF) |
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| ORF058388 | ABM | 1.0 ug DNA | EUR 506 |
Spock1 ORF Vector (Mouse) (pORF) |
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| ORF058389 | ABM | 1.0 ug DNA | EUR 506 |
Spock1 ORF Vector (Mouse) (pORF) |
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| ORF058390 | ABM | 1.0 ug DNA | EUR 506 |
Spock1 ORF Vector (Mouse) (pORF) |
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| ORF058391 | ABM | 1.0 ug DNA | EUR 506 |
 Anti-SPOCK1 antibody |
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| PAab08186 | Lifescience Market | 100 ug | EUR 386 |
 (pPB-C-His)) SPOCK1 Protein Vector (Human) (pPB-C-His) |
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| PV039937 | ABM | 500 ng | EUR 329 |
 (pPB-N-His)) SPOCK1 Protein Vector (Human) (pPB-N-His) |
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| PV039938 | ABM | 500 ng | EUR 329 |
 (pPM-C-HA)) SPOCK1 Protein Vector (Human) (pPM-C-HA) |
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| PV039939 | ABM | 500 ng | EUR 329 |
 (pPM-C-His)) SPOCK1 Protein Vector (Human) (pPM-C-His) |
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| PV039940 | ABM | 500 ng | EUR 329 |
 (OKBB01262)) SPOCK1 ELISA Kit (Human) (OKBB01262) |
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| OKBB01262 | Aviva Systems Biology | 96 Wells | EUR 505 |
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Description: Description of target: Testican-1 is a protein that in humans is encoded by the SPOCK1 gene. It is mapped to 5q31.2. This gene encodes the protein core of a seminal plasma proteoglycan containing chondroitin- and heparan-sulfate chains. The protein's function is unknown, although similarity to thyropin-type cysteine protease-inhibitors suggests its function may be related to protease inhibition. ;Species reactivity: Human;Application: ELISA;Assay info: ;Sensitivity: <10pg/ml |
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 SPOCK1 Antibody, HRP conjugated |
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| 1-CSB-PA730785LB01HU | Cusabio |
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Description: A polyclonal antibody against SPOCK1. Recognizes SPOCK1 from Human. This antibody is HRP conjugated. Tested in the following application: ELISA |
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 SPOCK1 Antibody, FITC conjugated |
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| 1-CSB-PA730785LC01HU | Cusabio |
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Description: A polyclonal antibody against SPOCK1. Recognizes SPOCK1 from Human. This antibody is FITC conjugated. Tested in the following application: ELISA |
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 SPOCK1 Antibody, Biotin conjugated |
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| 1-CSB-PA730785LD01HU | Cusabio |
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Description: A polyclonal antibody against SPOCK1. Recognizes SPOCK1 from Human. This antibody is Biotin conjugated. Tested in the following application: ELISA |
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) SPOCK1 Recombinant Protein (Mouse) |
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| RP175157 | ABM | 100 ug | Ask for price |
SPOCK1 Recombinant Protein (Mouse) |
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| RP175160 | ABM | 100 ug | Ask for price |
SPOCK1 Recombinant Protein (Mouse) |
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| RP175163 | ABM | 100 ug | Ask for price |
SPOCK1 Recombinant Protein (Mouse) |
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| RP175166 | ABM | 100 ug | Ask for price |
SPOCK1 Recombinant Protein (Mouse) |
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| RP175169 | ABM | 100 ug | Ask for price |
 Human Testican- 1, SPOCK1 ELISA KIT |
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| ELI-41909h | Lifescience Market | 96 Tests | EUR 824 |
) SPOCK1 sgRNA CRISPR Lentivector set (Human) |
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| K2278001 | ABM | 3 x 1.0 ug | EUR 339 |
 Recombinant Human Testican 1/SPOCK1 Protein |
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| RP01051 | Abclonal | 5 μg | EUR 136 |
 (pPB-C-His)) SPOCK1 Protein Vector (Mouse) (pPB-C-His) |
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| PV233546 | ABM | 500 ng | EUR 603 |
 (pPB-N-His)) SPOCK1 Protein Vector (Mouse) (pPB-N-His) |
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| PV233547 | ABM | 500 ng | EUR 603 |
 (pPM-C-HA)) SPOCK1 Protein Vector (Mouse) (pPM-C-HA) |
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| PV233548 | ABM | 500 ng | EUR 603 |
 (pPM-C-His)) SPOCK1 Protein Vector (Mouse) (pPM-C-His) |
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| PV233549 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPB-C-His) |
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| PV233550 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPB-N-His) |
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| PV233551 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPM-C-HA) |
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| PV233552 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPM-C-His) |
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| PV233553 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPB-C-His) |
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| PV233554 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPB-N-His) |
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| PV233555 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPM-C-HA) |
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| PV233556 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPM-C-His) |
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| PV233557 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPB-C-His) |
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| PV233558 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPB-N-His) |
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| PV233559 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPM-C-HA) |
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| PV233560 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPM-C-His) |
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| PV233561 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPB-C-His) |
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| PV233562 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPB-N-His) |
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| PV233563 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPM-C-HA) |
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| PV233564 | ABM | 500 ng | EUR 603 |
SPOCK1 Protein Vector (Mouse) (pPM-C-His) |
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| PV233565 | ABM | 500 ng | EUR 603 |
) Polyclonal SPOCK1 Antibody(N-term) |
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| APR13512G | Leading Biology | 0.1ml | EUR 528 |
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Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human SPOCK1 (N-term). This antibody is tested and proven to work in the following applications: |
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 ELISA Kit) Human Testican 1 / TIC1 (SPOCK1) ELISA Kit |
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| 20-abx258185 | Abbexa |
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 Human SPOCK1/Testican-1 PicoKine ELISA Kit |
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| EK1848 | BosterBio | 96 wells | EUR 425 |
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Description: For quantitative detection of human SPOCK1/Testican-1 in cell culture supernates, serum and plasma (heparin, EDTA). |
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 CLIA Kit) Human Testican 1 / TIC1 (SPOCK1) CLIA Kit |
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| 20-abx496043 | Abbexa |
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 (Target 1)) SPOCK1 sgRNA CRISPR Lentivector (Human) (Target 1) |
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| K2278002 | ABM | 1.0 ug DNA | EUR 154 |
 (Target 2)) SPOCK1 sgRNA CRISPR Lentivector (Human) (Target 2) |
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| K2278003 | ABM | 1.0 ug DNA | EUR 154 |
 (Target 3)) SPOCK1 sgRNA CRISPR Lentivector (Human) (Target 3) |
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| K2278004 | ABM | 1.0 ug DNA | EUR 154 |
) ELISA kit for Human Testican-1 (SPOCK1) |
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| KTE60335-48T | Abbkine | 48T | EUR 332 |
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Description: Quantitative sandwich ELISA for measuring Human Testican-1 (SPOCK1) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids. |
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ELISA kit for Human Testican-1 (SPOCK1) |
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| KTE60335-5platesof96wells | Abbkine | 5 plates of 96 wells | EUR 2115 |
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Description: Quantitative sandwich ELISA for measuring Human Testican-1 (SPOCK1) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids. |
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ELISA kit for Human Testican-1 (SPOCK1) |
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| KTE60335-96T | Abbkine | 96T | EUR 539 |
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Description: Quantitative sandwich ELISA for measuring Human Testican-1 (SPOCK1) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids. |
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 Mouse Testican- 1, Spock1 ELISA KIT |
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| ELI-41910m | Lifescience Market | 96 Tests | EUR 865 |
) Spock1 sgRNA CRISPR Lentivector set (Mouse) |
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| K3875701 | ABM | 3 x 1.0 ug | EUR 339 |
 (Target 1)) Spock1 sgRNA CRISPR Lentivector (Mouse) (Target 1) |
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| K3875702 | ABM | 1.0 ug DNA | EUR 154 |
 (Target 2)) Spock1 sgRNA CRISPR Lentivector (Mouse) (Target 2) |
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| K3875703 | ABM | 1.0 ug DNA | EUR 154 |
 (Target 3)) Spock1 sgRNA CRISPR Lentivector (Mouse) (Target 3) |
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| K3875704 | ABM | 1.0 ug DNA | EUR 154 |
) ELISA kit for Mouse Testican-1 (SPOCK1) |
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| KTE70235-48T | Abbkine | 48T | EUR 332 |
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Description: Quantitative sandwich ELISA for measuring Mouse Testican-1 (SPOCK1) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids. |
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ELISA kit for Mouse Testican-1 (SPOCK1) |
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| KTE70235-5platesof96wells | Abbkine | 5 plates of 96 wells | EUR 2115 |
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Description: Quantitative sandwich ELISA for measuring Mouse Testican-1 (SPOCK1) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids. |
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ELISA kit for Mouse Testican-1 (SPOCK1) |
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| KTE70235-96T | Abbkine | 96T | EUR 539 |
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Description: Quantitative sandwich ELISA for measuring Mouse Testican-1 (SPOCK1) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids. |
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 Spock1 3'UTR Luciferase Stable Cell Line |
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| TU119621 | ABM | 1.0 ml | Ask for price |
 Spock1 3'UTR GFP Stable Cell Line |
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| TU169621 | ABM | 1.0 ml | Ask for price |
Spock1 3'UTR Luciferase Stable Cell Line |
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| TU221107 | ABM | 1.0 ml | Ask for price |
 SPOCK1 3'UTR GFP Stable Cell Line |
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| TU074515 | ABM | 1.0 ml | EUR 2333 |
Spock1 3'UTR GFP Stable Cell Line |
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| TU271107 | ABM | 1.0 ml | Ask for price |
 SPOCK1 3'UTR Luciferase Stable Cell Line |
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| TU024515 | ABM | 1.0 ml | EUR 2333 |
) SPOCK1 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Human) |
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| K2278005 | ABM | 3 x 1.0 ug | EUR 376 |
 (Target 1)) SPOCK1 sgRNA CRISPR/Cas9 All-in-One Lentivector (Human) (Target 1) |
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| K2278006 | ABM | 1.0 ug DNA | EUR 167 |
 (Target 2)) SPOCK1 sgRNA CRISPR/Cas9 All-in-One Lentivector (Human) (Target 2) |
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| K2278007 | ABM | 1.0 ug DNA | EUR 167 |
 (Target 3)) SPOCK1 sgRNA CRISPR/Cas9 All-in-One Lentivector (Human) (Target 3) |
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| K2278008 | ABM | 1.0 ug DNA | EUR 167 |
) Spock1 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Mouse) |
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| K3875705 | ABM | 3 x 1.0 ug | EUR 376 |
 ELISA Kit) Human SPARC/Osteonectin, Cwcv And Kazal Like Domains Proteoglycan 1 (SPOCK1) ELISA Kit |
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| abx383426-96tests | Abbexa | 96 tests | EUR 911 |
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 Antibody) SPARC/Osteonectin, Cwcv And Kazal Like Domains Proteoglycan 1 (SPOCK1) Antibody |
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| abx145667-100ug | Abbexa | 100 ug | EUR 391 |
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SPARC/Osteonectin, Cwcv And Kazal Like Domains Proteoglycan 1 (SPOCK1) Antibody |
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| abx238186-100ug | Abbexa | 100 ug | EUR 509 |
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SPARC/Osteonectin, Cwcv And Kazal Like Domains Proteoglycan 1 (SPOCK1) Antibody |
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| 20-abx318007 | Abbexa |
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 (Target 1)) Spock1 sgRNA CRISPR/Cas9 All-in-One Lentivector (Mouse) (Target 1) |
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| K3875706 | ABM | 1.0 ug DNA | EUR 167 |
TRV-GFP: a modified Tobacco rattle virus vector for environment friendly and visualizable evaluation of gene operate.
Virus-induced gene silencing (VIGS) is a great tool for useful characterization of genes in crops. Sadly, the effectivity of an infection by Tobacco rattle virus (TRV) is comparatively low for some non-Solanaceae crops, that are economically necessary, comparable to rose (Rosa sp.).
Right here, to generate a simple traceable TRV vector, a inexperienced fluorescent protein (GFP) gene was tagged to the three’ terminus of the coat protein gene within the authentic TRV2 vector, and the silencing effectivity of the modified TRV-GFP vector was examined in a number of crops, together with Nicotiana benthamiana, Arabidopsis thaliana, rose, strawberry (Fragaria ananassa), and chrysanthemum (Dendranthema grandiflorum).
The outcomes confirmed that the effectivity of an infection by TRV-GFP was equal to that of the unique TRV vector in every examined plant. Unfold of the modified TRV virus was simple to observe by utilizing fluorescent microscopy and a hand-held UV lamp.
When TRV-GFP was used to silence the endogenous phytoene desaturase (PDS) gene in rose cuttings and seedlings, the standard photobleached phenotype was noticed in 75-80% crops which have been recognized as GFP optimistic by UV lamp.
As well as, the abundance of GFP protein, which represented the focus of TRV virus, was proved to correlate negatively with the extent of the PDS gene, suggesting that GFP might be used as an indicator of the diploma of silencing of a goal gene.
Taken collectively, this work gives a visualizable and environment friendly instrument to foretell optimistic gene silencing crops, which is effective for analysis into gene operate in crops, particularly for non-Solanaceae plant
A Mason-Pfizer Monkey virus Gag-GFP fusion vector permits visualization of capsid transport in dwell cells and demonstrates a job for microtubules.
Immature capsids of the Betaretrovirus, Mason-Pfizer Monkey virus (M-PMV), are assembled within the pericentriolar area of the cell, and are then transported to the plasma membrane for budding.
Though a number of research, using mutagenesis, biochemistry, and immunofluorescence, have outlined the position of some viral and host cells elements concerned in these processes, they’ve the drawback of inhabitants evaluation, relatively than analyzing particular person capsid motion in actual time.
On this examine, we created an M-PMV vector wherein the improved inexperienced fluorescent protein, eGFP, was fused to the carboxyl-terminus of the M-PMV Gag polyprotein, to create a Gag-GFP fusion that might be visualized in dwell cells.
In an effort to categorical this fusion protein within the context of an M-PMV proviral spine, it was essential to codon-optimize gag, optimize the Kozak sequence previous the initiating methionine, and mutate an inside methionine codon to 1 for alanine (M100A) to forestall inside initiation of translation.
Co-expression of this pSARM-Gag-GFP-M100A vector with a WT M-PMV provirus resulted in environment friendly meeting and launch of capsids.
Outcomes from fixed-cell immunofluorescence and pulse-chase analyses of untamed sort and mutant Gag-GFP constructs demonstrated comparable intracellular localization and launch of capsids to untagged counterparts. Actual-time, live-cell visualization and evaluation of the GFP-tagged capsids supplied robust proof for a job for microtubules within the intracellular transport of M-PMV capsids. Thus, this M-PMV Gag-GFP vector is a great tool for figuring out novel virus-cell interactions concerned in intracellular M-PMV capsid transport in a dynamic, real-time system
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