Service
- Multi-Plex Immunoassay (MPI)

Introduction

The major activity of this core is to provide multi-plex assay to measure mediator levels in patients’ sera.

Multi-Plex Immunoassay (MPI) is a technology that allows for the simultaneous quantification of analytes in serum/plasma and culture supernatants, making them a useful tool in both disease mechanistic studies and in biomarker discovery. We currently offer the analysis of cytokines and chemokines pertaining to immunology and our detection panels are fully validated. MPI panels are also customizable to the investigators’ needs. For projects, prior consultation with the laboratory is recommended to ensure optimal service information.

The multiplex is capable of measuring up to 100 analytes. One reaction consists of magnetic COOH beads dyed with different concentrations of fluorophores to generate distinct bead sets. Each bead set is coated with a capture antibody specific for a specific cytokine. A second biotinylated detection antibody and streptavidin-phycoerythrin (S-PE) is used to detect the captured cytokine. The Bio-plex analyzer is a dual laser, flow-based, sorting and detection platform. One laser detects the beads and determines which cytokine is being detected. The second laser determines the magnitude of PE-derived signal, which is labelled on the detection antibody and is proportional to the amount of cytokine bound.


Process

process

Sample Preparation

Serum prep

  • Blood is recommended to prepare in the blood collection tubes with YELLOW cover (BD MICROTAINER. SSTTM)
  • If blood is not prepared in the blood collection tube with yellow cover, allow blood to clot at room temperature for 30-45 mins before proceeding to centrifugation steps (not recommend-high variation).
  • Centrifuge at 1,000 x g for 15 mins at 4℃ and transfer into a clean tube.
  • Repeat centrifuge at 1,000 x g for 10 mins at 4℃ to completely remove platelets and precipitates.
  • Aliquot and store undiluted sample in 1.5 mL eppendorf at -80 ℃ if not using immediately.

Plasma prep

  • Use blood collection tubes with PURPLE cover (BD MICROTAINER. SSTTM) or collection tubes coated with anticoagulant (EDTA and Heparin are the recommended anticoagulant for preparing plasma samples)
  • Centrifuge at 1,000 x g for 15 mins at 4℃ and transfer into a clean tube.
  • Repeat centrifuge at 1,000 x g for 10 mins at 4℃ to completely remove platelets and precipitates.
  • Aliquot and store undiluted sample at -80℃ if not using immediately.

Volume of sample required

  • For serum and plasma samples, we request 300 µL of human sample and 150 µL of mouse sample.
  • For tissue lysate samples, we request 150 µL of 0.8 mg/mL sample.
  • For culture medium samples, please prepare extra 15 mL culture medium for us as standard curve buffer.
  • If the mediator number is over 24, please contact the ICF to provide more sample volume.

Note

  • For the cytokine Analysis, each 96-well plate can accommodate 80 unknowns. 16 wells are required for the cytokine standards and blanks. We highly recommended that the entire sample will be run in duplicates (40 unknowns/plate).
  • Please DO NOT SEND infectious, or potentially infectious, human samples.
  • Please use protease inhibitor when profiling for diabetes/metabolic markers.
  • Extremely lipemic samples may be filtered with a 0.22 µm filter to prevent clogging.
  • Hemolyzed samples may not be suitable for Multi-Plex Immunoassays.
  • Serum is qualitatively different from plasma because of how the removal of fibrinogen and other proteins that form fibrin clots affects coagulation.
  • Protein concentration of serum is 3~4% less than that of plasma.

Shipping and Contact information

  • Please ship/deliver all the samples at -70 ℃
  • Delivery address: Inflammation Core Facility (Room N431), Institute of Biomedical Sciences, Academia Sinica., No. 128, Sec. 2, Academia Rd, Nankang Dist., Taipei City 115, Taiwan (ROC)
  • Recipient : Dr. Huan-Yuan, Chen (陳煥源)
  • Tel: +886-2-2652-3945 (Lab)

Assay Range

MPI assay range for human mediators

MPI assay range for human mediators

MPI assay range for mouse mediators

MPI assay range for mouse mediators

Price

For detecting 6 mediators in one 96 well plate

Research institute (NTD/plate) Company (NTD/plate)
33,000 70,000

For detecting more mediators (N≧6) in one 96 well plate

Research institute (NTD/plate) Company (NTD/plate)
33,000 x N/6 70,000 x N/6

Note: Technically, 80 unknowns can be assayed in one plate. We recommend users apply duplicated assay (40 unknowns/ plate).


Payment Procedure

billing procedures pic

Payment Procedure

  1. Users fill in application form.
  2. After the application forms are approved, the facility will send a remittance advice email to each applicant.
  3. Arrange the payment.
  4. After the payment has arrived, the receipt and application form will be sent to users
  5. Upon receiving the sample delivery notification and the receipt, you may send the samples to the facility during working hours.

※A partial payment is also accepted if the total payment exceeds your quota. (For example, if the estimated price exceeds 300,000 NTD, two payments of 50% each can be made on the current and following month)

[Notice for payment]
  1. The charge is non-refundable, so please confirm the service charge before payment. After the payment has arrived, we will start to execute the service.
  2. If you have some special reasons for not delivering the payment before the service , please fill the “Deferred Payment Affidavit Letter”( Download here ) and send it to the core. The core manager will confirm this letter and execute this service before the payment. Each case can only apply once.
  3. Due to the different purchasing process of each administrative unit, it may takes 3-4 weeks from user submitting the purchase request to the unit to they actually allocate the funds to the Academia Sinica account. Please take the time requirement into consideration during scheduling the experiment.
[Notice for sample delivery]
  1. After the core ahs received the payment, we will send a “Sample Delivery Notification” to user. Please deliver your sample within one month. If we don’t receive the sample within a month, we will send the sample delivery notification again. However, if the sample is not delivered for more than three months, the case will be regarded as abandonment and no re-fund will be given.
  2. If there are some special circumstances that the user need to extend the delivery deadline, please inform the core and tell us the expected sample delivery date. If the expected delivery date is not clearly informed, the delivery deadline will be automatically extended by one month. Each case can only apply once.

Payment method for users in Academia Sinica

  • Please reply the email to agree the fund transfer.
  • The facility will transfer the payment through the in-house PI payment system (quota).
  • If users in Academia Sinica use Ministry of Science and Technology (MOST) funds to pay for the use of facilities, the charge calculation will be based on the "Research institute outside Academia Sinica" fee standard

Payment methods for users in other academia units or company

Please choose one payment method from below.

  1. Wire transfer
    • Beneficiary account number: 004056030013
    • Beneficiary account name: 中央研究院301專戶
    • Beneficiary bank and branch: LAND BANK OF TAIWAN NAN KANG BRANCH.
    • Please provide the information below via Email/fax after remit the money.
      1. Name of core facility service you use
      2. Client’s Title
      3. Client’s Name
      4. Client’s Address
      5. Amount of receipts (one receipt for each case/or one receipt for multiple cases)
      6. Confirmation of the payment (wire transfer/ cash remittance note)
      7. Service application form
    • Receipt will be sent within one week
  2. Send a check by registered mail
    • Check title: Academia Sinica (中央研究院)
    • Mail address: Room 2103, Academic Affairs and Instrument Service, Academia Sinica., No. 128, Sec. 2, Academia Rd, Nankang Dist., Taipei City 11529, Taiwan (ROC)
    • Recipient: Mr. Wun-Ci Ciou (邱文棋)
    • Receipt will be sent within one week
  3. Direct cash/ check deposit
    • Please bring cash or check to Room 2103, Administrative Building, Instrument Management Department, Academic Affairs and Instrument Service, Academia Sinica and look for Mr. Wun-Ci Ciou (邱文棋)
    • The receipt will be given to you at once

Order

  • Please submit your application form to: inflamcore@ibms.sinica.edu.tw
  • Multi-Plex Immunoassay Set:
    • Human 6-Plex: IL-1β, IL-6, IL-8, IL-10, IL-17A, TNF-α
    • Human 12-Plex: IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12(p70) , IL-13, IL-15, IL-17A, TNF-α, IFN-γ
    • Mouse 6-Plex: IL-1β, IL-6, IL-10, IL-17A, TNF-α, IFN-γ
    • Mouse 12-Plex: IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, IL-17A, IL-23, TNF-α, IFN-γ
  • ICF offer customization services which clients can choose the mediators they want to detect.

Download form

order pic1
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