Antagene, Inc. was founded in the Bay area of San Francisco, California. The principal goal of the company is to provide products and services that ultimately support academic and commercial endeavors in the area of :
We achieve this goal through our commitment to offer the most characterized, highest quality service and products available today based on our innovative technologies and our enormous research and development capabilities.
In pursuit to offer the top-quality products, we utilize the most technologically advanced equipments to thoroughly test our products. Our product data will include the most current ELISA, High Performance Liquid Chromatography (HPLC), Mass Spectro, SDS-PAGE, Western blotting, Immuno-histochemistry data for the most stringent QA and QC inspections.
Antagene Inc. is ready to commit its skills, laboratories, and equipment to every client's research project. Our customer service team provides flexible and multiple choices to meet the unique requirement of each of our clients. Our goals are to continue providing the best quality of service and products to today's scientific research community.
For many years, immunoassays/diagnostic kits have been used in hospitals, laboratory medicine, and research to identify and assess the progression of disease. Information gained by clinical immunoassay testing has led to improved therapeutic choices. In research laboratories, immunoassays are used in the study of biological systems by tracking different proteins, hormones, and antibodies. In industry, immunoassays are used to detect contaminants in food and water. Antagene Inc is a custom service company specializing in antibody production, immunoassay development and kit manufacturing. The scientists at Antagene Inc have the knowledge and expertise to develop reliable, sensitive, and specific immunoassays. Antagene Inc can develop immunoassays:
for immunogenicity studies to detect IgG, IgE, IgM, and other molecules to measure the immune response to a specific drug or targeted proteins. to measure proteins and peptides found in serum, plasma, urine, and cell culture fluids for research or diagnostic purpose. to measure enzymes used in food processing and industrial applications for precise quality control.
If you need to detect a specific target protein or drug, we can fully develop immunoassay test (or optimize an existing assay) according to your specifications. Once developed, we will ship the components to you, and all products associated with the project shall become the exclusive property of yours at the conclusion of the project. Full confidentiality is guaranteed. For more details regarding our bioassay and kit development services, please contact us.
Antagene offers ascites production from small Research & Development scale to large-scale production. We secured a USDA registered research facility, and we comply with all federal guidelines for care and use of laboratory animals.
Priming and cell preparation:
You may provide sufficient cells from designated cell line (approximately 2-2.5 x 106 cells per animal). A minimum of 5 mice will be used per hybridoma clone. As soon as we receive your order, we will start to prime at least 10 mice per hybridoma clone with Pristane for you. (Default: minimum 14 days of priming, with 0.5 ml of Pristane / mouse). The time interval between priming and inoculation of hybridoma cells is 10-14 days. Generally, very high concentrations of cells are associated with greater mortality and concentrations < 1 x 105 cells elicit fewer ascitic tumors and these tend to have a smaller volume yield. Cell suspensions should be prepared sterilely in physiological solutions.
If you prefer to send us frozen vials, 3 vials of cells (3x106 cells / vial) from designated cell line are required. As soon as we received the sample, Cells are thawed and viability is tested. We then begin to expand cells to appropriate number for inoculation (Default: 2-3 X 106 cells/ animal). We determine percent viability via Trypan Blue Dye. Approximately one or two weeks are needed to prepare cells from frozen stocks for injection, and there is an additional charge ($300/clone) for this service.
(Additional: Nude and SCID mice are available for ascites production for non-murine hybridoma cell lines.)
Harvesting Ascites Fluid:
Hybridomas are injected into pristane-primed mice, then mice are monitored daily for ascites development and general condition. Ascites collection begins when accumulation of ascites becomes evident and continues until final harvests are performed. You can choose to receive unpurified, defibrinated ascites, or Antagene will purify ascites for you. In general, ascites production is completed within 2-3 weeks after injection of hybridomas.
The ascites fluid will be sent back to you via overnight transporter. If you ask for antibody purification, we will use those ascites to purify antibody (additional charge $500 for IgG purification), 15-20mg purified antibodies will be delivered at the end. Shipment costs to your laboratory are additional as indicated in our invoice.
We will keep no sample after the experiment achievement. No sample will be transferred to any third party or be commercially used. You will remain the only proprietary of the hybridoma. Moreover, we will not disclose to any third party that we have used your material for any manipulation or worked with your company. Starting with your own hybridoma, we cannot guarantee ascites quality in case of low antibody yield.
We are pleased to offer experience in the design, synthesis and production of peptides using t-Boc/Fmoc solid phase and solution phase technology. We produce high quality peptides with a fast turnaround time.
For multigram synthesis (or large scale synthesis) project, please contact us for a quick and convenient peptide quote.
Phase I: Peptide synthesis (2 weeks)
Peptide synthesis up to 15 amino acids.
Each phospho-residue (Phospho-Ser, Tyr or Thr)
Phase II. Conjugation (1 week)
Conjugation of 5 mg peptide (using Cysteine) to KLH.
10 mg free peptide will be provided for client
Phase III. Immunization and anti-sera production (10 weeks)
Ten-week protocol to produce antibodies in 2 rabbits.
Pre-immune 5 ml/per rabbit and total final bleeds from two rabbits (100 ml antiserum) will be provided to client.
IV. Additional Bleed-Out Service (1 week)
provide 50 ml from each rabbit
V. Re-boosting Immunizations (7 weeks)
(two additional booster injection, 20 days interval, 50 ml antiserum bleed after 50 days)
VI. ELISA Test (1 week)
ELISA test for final total bleed (including conjugating peptide to BSA)
VII. Purification (1 week)
Synthesis the control peptide of the same sequence without phospho residue.
Phospho-peptide antibody purification by 2-steps peptide affinity purification including phospho-peptide column and non-phospho-peptide column
ELISA data will be included for free
Phosphospecific Antibody Affinity Purification
Antagene Inc. offers phosphospecific antibody affinity purification including:
Antagene Inc will provide the customer with the service in the shortest possible time. Delays may be incurred if the antigen is poorly immunogenic .
All the products of this project shall become the exclusive property of the client at the conclusion of the project. Full confidentiality is guaranteed.
Phase 1: Immunization (8-10 weeks)
Immunization of Balb/c mice (up to 5 animals) with antigen to develop antiserums.
The client may choose to follow our standard immunization protocol or to use your own protocol. In most cases, we injected the mouse every two weeks for a total of 6 weeks before doing a test bleed on the mouse. The test bleed samples are then examined for antibody production using ELISA (The client may request up to 0.5 ml serum sample/per mouse for their own additional characterization). ELISA will be used to determine the highest titer mice to be used for cell fusion. The immunization process may be extended if the test result is not optimal.
Antigen requirement: 0.5 mg of purified recombinant protein or intact protein will be required for immunization.
With peptide synthesis request: Custom Peptide synthesis up to 15 amino acids (additional amino acid for $12 per residue) with HPLC purification up to 90% purity, peptide KLH conjugation for immunization, peptide BSA conjugation for screen. 10 mg free peptide will be available for the client.
Phase 2: Fusion (2-6 weeks)
Fusion of spleen cells with myeloma cell line. The fused cells are then placed in 96-well culture plates. Viable colonies appear as early as 3-5 days after fusion and are kept healthy with a fresh supply of nutrient media as necessary.
Phase 3: Screening 2-6 weeks
When hybridoma colonies are large enough to screen, screening of all wells containing fusion products is done with ELISA assay. All positive wells will be expanded and frozen for parental hybridomas. The number of positives depends on the nature of the antigen, and can range from a few hybridomas to a dozen or more. Additional hybrids can be frozen for further studies.
Phase 4: Stabilization and Expension of clones 2-4 weeks
After positive hybridomas are selected for further development, they will be cloned using the limited dilution method and screened by ELISA. 5 clones with the desired specificity will be expanded, and hybridoma medium (50 - 100 ml) will be collected for further studies
Purification 1 week
The secreted antibody will be purified by Protein G affinity chromatography and deliver to the customer (Approximately 20-30 mg).
Antibodies are eluted using both pH11 and pH2.5 buffers and neutralized immediately followed by dialysis in PBS
Phase I. Peptide Design and Synthesis (2 weeks)
We start with the synthesis of custom peptide for up to 15 amino acids. ($12 for each additional residue).Special Assistants are provided for peptide design free of charge.
Phase II. Conjugation (1 week)
ï¿½ It takes 1 week to conjugate 10 mg of peptide to KLH (using Cysteine).
ï¿½ We will provide 10 mg of peptide to the client for free.
Phase III. Immunization & Anti-Sera Production (10 weeks)
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