Mammalian-expressed proteins are important bioreagents to accelerate your research. Abnova provides a full-service mammalian protein encompassing cloning, expression, production, and purification based on proprietary expression plasmids, His8-Sumo and IgG1 Fc tags, PEI transfection in HEK293 and CHO cells, and Ni-NTA or Protein A affinity column purification. The purified protein is checked by SDS-PAGE and Western Blot. Customers can also provide downstream assays to confirm the bioactivity of the protein. Our technical support is available to accommodate additional requests and needs required by your experiments. Abnova takes care of the proteins so you can focus on your research! We guarantee the bioactivity of our custom mammalian-expressed proteins.

Flowchart

Mammalian_Protein_Flowchart.jpg (79 KB)

 
 

Specification

Mammalian Transient Protein Expression Service Details

Expression vector

His8-Sumo tag at the

N-terminus

Human lgG1 Fc tag at the

C-terminus

  Timeline
 Cell line  HEK 293  CHO  HEK 293  CHO

Phase I

Cloning of your gene of interest into a vector with His8-Sumo tag 

at the N-terminus or human lgG1-Fc tag 

4 weeks

Phase II

50 mL transient expression in mammalian cells using polyethyleneimine (PEI)

6 weeks

  

Phase III

Protein purification from 50 mL of HEK 293 cell culture using Ni-NTA affinity purification columns

Protein purification from 50 mL of CHO cell culture using Ni-NTA affinity purification columns

Protein purification from 50 mL of HEK 293 cell culture using Protein A affinity purification columns

Protein purification from 50 mL of CHO cell culture using Protein A affinity purification columns

Deliverables

• Purified protein   • SDS-PAGE report   • Western blotting   • CoA

Advantages
  • High efficiency
  • Optimized mammalian expression system
  • High-throughput and large-scale production
  • High yield expression with post-translational modification and correct folding
 
 

Examples

ELISA Results of Human IL-12 Biological Activity

 
Measure by its ability to induce IFN gamma secretion in PHA-activated human peripheral blood lymphocytes (PBMC). The ED50 for this effect is 0.05~0.2 ng/mL.
 

ELISA Results of HMGB1 Biological Activity

 

(A) Human HMGB1 recombinant protein induced TNF alpha with a dose-dependent manner in RAW264.7 cells.
(B) Human HMGB1 C23AC45A mutated recombinant protein induced TNF alpha with a lower amount than wild-type HMGB1 protein in RAW264.7 cells.
(C) Human HMGB1 C23AC45AC106A mutated recombinant protein almost not induce TNF alpha in RAW264.7 cells.
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