pFUSE-Lucia-CHIg-mG2b
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Cat.code:
pfuselc-mchg2b
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ABOUT
Heavy chain constant region expression plasmid - Mouse IgG2b - Mouse IgG2b tag
pFUSE-Lucia-CHIg-mG2b is a cloning plasmid that expresses the constant region of the murine IgG2b heavy chain. It contains a multiple cloning site upstream of the constant region to enable cloning of the heavy chain variable region.
This plasmid is designed for antibody generation when co-transfected into mammalian cells with the light chain variable region cloned into pFUSE-CLIg.
pFUSE-Lucia-CHIg-mG2b contain the secreted luciferase (Lucia) gene upstream of the MCS and the CH region to serve as a tag to facilitate the detection and quantification of recombinant antibodies.
Specifications
- Constant region of the mouse IgG2b heavy chain
- Lucia secreted luciferase tag
Lucia luciferase is a secreted protein, therefore, the VH sequence should not include a signal sequence.
Plasmids is selectable with Zeocin™ in E.coli and mammalian cells.
These products are covered by a Limited Use License (See Terms and Conditions).
All products are for research use only, and not for human or veterinary use.
Contents
- 20 µg of lyophilized DNA
- 1 ml of Zeocin® (100 mg/ml)
Product is shipped at room temperature
SPECIFICATIONS
Specifications
In vitro transfection, antibody generation
In vitro transfection
Plasmid construct is confirmed by restriction analysis and full-length open reading frame (ORF) sequencing.
CONTENTS
Contents
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Product:pFUSE-Lucia-CHIg-mG2b
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Cat code:pfuselc-mchg2b
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Quantity:20 µg
1 ml of Zeocin® (100 mg/ml)
Shipping & Storage
- Shipping method: Room temperature
- Upon receipt, store product at -20 °C.
Storage:
Details
1- Schematic representation of a Lucia luciferase-tagged antibody
2- Luciferase activity of Lucia-tagged anti-hTNF-α antibodies.
CHO cells were stably co-transfected with a heavy chain expressing plasmid, pFUSE-Lucia-TNF-CHIg-hA2m1 (TNF-CHIg-hA2m1) or pFUSE-Lucia-TNF-CHIg-hG1 (TNF-CHIg-hG1) and a light chain expressing plasmid, pFUSE2-TNF-CLIg-hk (TNF-CLIg-hk) to generate Lucia-tagged anti-hTNF-α antibodies, Lucia-anti-hTNF-α-hIgA2m1and Lucia-anti-hTNF-α-hIgG1, of human IgAm2 and human IgG1 isotypes, respectively. pFUSE2-CLIg-hk (CLIg-hk), which expresses no VL, and pSELECT-blasti-mcs (mcs) were used as negative controls. Supernatants
were collected and the luciferase levels determined using QUANTI-Luc™. Only the cells co-producing a Lucia-anti-hTNF-α heavy-chain and an anti-hTNF-α light chain displayed luciferase activity.
3- Neutralizing activity of anti-hTNF-a antibodies
The activity of anti-hTNF-α-hIgA2m1 and Lucia-anti-hTNF-α-hIgA2m1 antibodies, purified from the supernatants of CHO transfected cells, was determined by performing an TNF-α neutralizing assay. Both antibodies display similar neutralizing activities, thus fusion of the Lucia luciferase tag at the N-terminus of the heavy chain does not alter the functionality of the antibody.
DOCUMENTS
Documents
Technical Data Sheet
Safety Data Sheet
Plasmid Sequence
Certificate of analysis
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