Catalogue Number: 41115-1-PBL
For over 30 years, PBL Assay Science has had a mission to help ‘solve the most difficult problems’ of researchers around the world. From its founding in 1990 and continuing through today, PBL has become widely known as a source for high-quality immunoassay kits, cytokines and interferons, monoclonal and polyclonal antibodies, and other related reagents for life science researchers. PBL products can provide reliable and consistent solutions for all your Assay Science needs.
Singleplex ELISAs: PBL’s ELISA kits are a comprehensive range of heterogeneous antibody sandwich immunoassays. The detection system utilizes horseradish peroxidase (HRP) and Tetramethyl-benzidine (TMB) as the enzyme-substrate which provides sensitive and robust assays for the detection and quantification of interferons and cytokines.
Multiplex ELISAs: Fully quantitative high throughput multiplex ELISA kits that provide simultaneous quantification and detection of up to 16 different analytes from a single sample.
Interferons: PBL offers a wide range of interferons from different species and of different classes including multiple IFN-alpha and IFN-beta subtypes. Our interferons are high purity, provide consistent performance and biological activity and are suitable for use in a range of research applications.
Cytokines: Cytokines are a class of signaling proteins that play a role in both the induction and effector phases of all immune and inflammatory responses, including cell proliferation, intercellular communication, and cell death. Cytokines are produced by a variety of hematopoietic and non-hematopoietic cell types and can exert autocrine and paracrine effects. They are related more correctly to hormones than to growth factors in their overall functions, but many cytokines also exhibit growth factor activity.
Monoclonal and Polyclonal Antibodies: PBL offers a range of both monoclonal and polyclonal antibodies raised against different interferons (IFNs) from various mammalian species. As opposed to other commercially available Anti-IFN antibodies, PBL’s human IFN alpha antibodies are tested against a wide range of human IFN subtypes
At PBL, we understand the difficulties involved in working with a partner outside of your organization—because it’s something we’ve been doing for 30 years. Whether you have limited internal capacity or whether you’re seeking outside expertise in addressing specific experimental challenges, we know it takes an investment of your time and energy to establish a relationship and bring your projects to completion. But there’s one thing we do know, that’s been proven time and time again: PBL Assay Science is a partner that can take a great burden off your shoulders. We take the time to listen, to understand what you want to accomplish; we provide guidance from our deep knowledge base; we recommend only the work that your research requires; and we do all of this under the utmost confidentiality.
We don’t just run immunoassays and spit out results—we can be your ‘Thought Partner’. We can do the work. We can deliver results. Contact Us today to discuss your research needs. For simplified sourcing, we also work with Science Exchange and Scientist.com.
Here are some of the services we provide and an analyte selector tool to view immunoassay services available from PBL on different platforms:
Find out more in the sections below:
PBL can expedite your R&D work as our technologies and services can be tailored to meet your specific requirements under fit-for-purpose guidelines. For many of your sample testing and screening needs, PBL can help you measure your analyte(s) of interest. From state-of-the-art technologies quantifying into the femtogram/milliliter level to simultaneous detection of up to 9 analytes in a single well, PBL offers a multitude of options to fit your project needs.
We understand the value of your experimental samples and execute assays just as you would – consuming a minimum of sample while generating robust results. We can take on work you don’t have the capacity to handle yourself. Let us help you develop, customize, or just execute assays on your samples in a timely, professional manner.
High precision. Unprecedented sensitivity. Digital readout. Take your detection to the next level.
The unique immunoassay technology provided by the Simoa® platform allows PBL to deliver fg/ml level sensitivity for low abundance biomarkers in serum, plasma, and other matrices. Single Molecule Array technology traps a single bead in a femtoliter-sized well allowing for “digital” measurement of each bead. This approach can increase assay sensitivities several orders of magnitude over conventional bioassays.
The combination of this instrument’s markedly heightened sensitivity and its’ capacity for digital analyte measurement ensures accuracy and precision across a wide dynamic range and minimizes sample volume requirements.
Figure 1. Quanterix Simoa® Technology Overview
Unprecedented precision can be obtained using Simoa’s “digital” readings of single molecular events. In the image above, individual paramagnetic beads are trapped in femtoliter-sized wells, then sealed over and quantified by SIMOA image analysis.
Accurate and robust measurement. Increase your understanding. Detect at unprecedented levels.
The ability to accurately measure low-abundance analytes present in complex matrices is essential for the profiling and characterization of cytokines and other biomarker analytes. Employing proprietary single molecule counting technology with robust microparticle-based SMCxPRO® & Erenna® immunoassays, PBL’s cytokine detection services can provide scientists with sub-pg/ml level measurements of low-abundance analytes in healthy or disease sera and plasma.
An increased understanding of the role and regulation of cytokines in disease states results from greater profiling and characterization of their activities in biological responses. Singulex technology can be instrumental in furthering our collective understanding of low-abundance cytokines in complex matrices.
Figure 2. EMD Millipore Single Molecule Counting Technology Overview
Low abundance biomarkers in complex biological samples are transferred to a 96-well plate and incubated with capture antibody-coated microparticles. After washing and incubating with dye-labeled detection antibodes, wells are again washed to remove unbound detection antibodies. An elution buffer is added, and the eluate drawn into the instrument for single-molecule counting.
Broad dynamic range. Minimal background. High sensitivity. Enhanced signal capture achieved through luminescent detection.
PBL offers high precision protein quantification services for biomarker detection in a broad range of sample matrices for Human, Non-Human Primate (NHP), Mouse, Rat, and Canine samples. PBL utilizes MSD’s validated assay kits (V-Plex) to quantify up to 54 analytes within a single sample. This enables simultaneous comparison of the expression of pertinent cytokines, chemokines, and other biomarkers in samples from diseased and normal patients at different intervals.
Multiplex immunoassays on the MSD platform maintain the sensitivity and performance offered by singleplex ELISAs while providing additional benefits such as cost-savings and targeting of several related analytes in a single sample. The ability to quantify compatible analytes while requiring no more than 25 μl of neat sample ensures efficient use of precious sample.
Figure 3. MSD-ECL Technology Overview
Unique detection technology utilizes SULFO-TAGTM labels which emit light upon electrochemical stimulation. The intensity of light generated is captured via CCD camera and pixel intensities quantified. This system provides the basis for achieving measurement of a several log range of biomarker expression levels in a variety of matrices.
Sensitive detection and robust results. Dedicated, flexible workforce. Reliable, high-quality outcomes.
Take advantage of PBL ELISAs or other commercial ELISAs for a wide range of additional analytes using PBL’s services.
PBL ELISA kits are heterogeneous antibody sandwich immunoassays. The antigen is captured by an antibody immobilized to the wells in a microtiter plate and a secondary antibody is used to complete the sandwich. The detection system which utilizes horseradish peroxidase (HRP) and tetramethyl-benzidine (TMB) as the enzyme-substrate provides a sensitive and robust assay for the detection and quantification of interferons and cytokines.
PBL routinely runs ELISA studies for our clients for the detection and differentiation of interferons and cytokines in various sample matrices. We offer a range of heterogeneous immunoassay services for:
High sensitivity & specificity using low sample volumes. Cost-effective quantitation of multiple analytes.
PBL’s cost-effective multiplex ELISA services enable simultaneous detection of up to 16 different analytes in a single sample.
These ELISAs provide chemiluminescent results on each analyte and allow for a global understanding of ongoing immune responses in diseases ranging from cancer to autoimmunity. Evaluation of one given inflammatory molecule in the context of several others, repeated measurements of the same cytokine panels in the same subjects under different treatment assay conditions, and reliable detection of different proteins across a broad range of concentrations are just some of the benefits provided by this multiplex service.
Figure 4. Multiplex Technology Overview
PBL’s multiplex ELISAs employ a technology in which distinct capture antibodies are individually spotted into each well of a 96-well ELISA plate in a defined array. These plates are run similar to a normal ELISA run. An image of the plate is then taken using a CCD camera.
Using image analysis software to perform subsequent analysis, the chemiluminescent results for each analyte allow for a deeper understanding of the cytokine profiles of the samples.
PBL’s reagent development services offer customized production of cytokines, interferons, antibodies, and other proteins.
With a wealth of knowledge on immunological interactions, we offer protein biophysical characterization services which can determine the kinetic characterization of protein-protein interactions. These services include determination of:
PBL offers biophysical characterization of antibody affinity for a ligand or the measurement of antigen:antibody Kon/Koff rates. To accelerate assay development, we also determine which antibodies bind to antigens in a sandwich format. Binning— determination of antibodies which bind to non-overlapping epitopes—is performed using multiple hybridoma clones.
The graph below shows the association and dissociation of IFN to a surface-immobilized antibody as determined by real time Biolayer Interferometry (BLI) using varying concentrations of IFN. The raw data is shown in blue and the model response in red.
Figure 1. IFN Binding to and Dissociation from a Surface-Immobilized Monoclonal Antibody Evaluated by Bio-Layer Interferometry (BLI)
PBL offers custom antibody labeling using a range of dyes and labels including FITC, R-PE, biotin, etc.
The graph below shows an example of a R-Phycoerythrin (PE) Conjugation of a Mouse Monoclonal Antibody.
Figure 2. Flow cytometry analysis of KG-1 cells showed expression of Human Interferon Alpha/Beta Receptor. Isotype control (shaded histogram) and human interferon alpha/beta receptor (un-shaded histogram). (Catalog No. 21370-3)
Cell-based assay tools are key to understanding cellular mechanisms in a biologically relevant context. PBL offers a targeted portfolio of cell-based assay services to provide you with the most relevant information and facilitate advancement of your research.
Whatever your cell-based study needs, PBL will work with you to determine the most suitable assay program to meet your goals. PBL’s services team is a trusted partner of midsize and major biotech and pharma companies worldwide.
Growth promotion and inhibition assays are often used to measure cytokine and drug activity profiles. Such Proliferation and Anti-Proliferation (AP) assays utilize a variety of cell lines and protein or small molecule stimuli to address specific applications and clinical indications. EC50 or IC50 results are provided.
Figure 1 shows OVCAR-3 grown in the presence of IFN-alpha. Cellular anti-proliferation was quantified using soluble tetrazolium reagent. Figure 2 shows TF-1 grown in the presence of GM-CSF. Cellular proliferation was quantified using soluble tetrazolium reagent. For both graphs, the quantity of reduced reagent product is directly proportional to viable cell number.
Figures 1 and 2. Growth Inhibition and Promotion Assays
Accurate measurement of the ability of a sample to induce cytokine production and/or release from cell lines or isolated donor cells is the standard assay for certain applications, such as measurement of IL-18 activity.
Figure 3 shows a natural killer cell model, NK-92, stimulated to produce IFN-Gamma by incubation with Type I IFNs. The data is graphed as a function of IFN-alpha concentration in the bioassay.
Figure 3. Cytokine Secretion Assay
The standard bioassay used to determine the biological activity of interferon (IFN) encompasses measurement of the protection of cells from the cytopathic effect (CPE) of certain viruses. CPE assay services may be used to measure the antiviral activity of IFN-α, β, ω, γ, and λ in human samples, as well as in a variety of other species, and in multiple matrices.
CPE assays are inherently complex due to the metabolic state of cells, virus replication, and the ability of IFN to protect cells. Our years of knowledge and expertise in CPE assays will reduce variability and provide you with reproducible quality results.
Results are provided either as a graphical representation of dye binding which allows slope and parallel line analysis, or visually determined from microscopic examination of the CPE and determination of the dilution of test samples which protects 50% of the cells.
Figure 4 to the right shows IFN-beta titrated in a CPE assay on A549 cells with EMCV. In this antiviral assay, the EC50 is approximately 1 unit/ml of interferon. Units of IFN activity were determined colorimetrically with respect to an international reference standard for human IFN.
Figure 4. Human IFN-Beta AVA Standard in CPE Assay
The standard method of accurately determining whether a sample may inhibit the biological activity of cytokines is the neutralization assay. Results, provided either as a visual read or in a graphical format, reveal the ability of a sample to block the activity of IFN added at a specific concentration which provides 100% cell protection.
May be used as an anti-drug antibody (ADA) assay for certain therapeutics.
Figure 5 to the right shows representative neutralization curves of two different MAbs to IFN in a CPE assay to determine neutralization titer. One neutralization unit (NU) is the amount of antiserum or antibody (NAb) required to neutralize 1 unit of human IFN to a 50% endpoint. NU are determined with respect to the international reference standard.
Figure 5. Monoclonal Antibody Titration (Neutralizing Antibody Assay)
As the drug discovery pathway shifts toward specifically targeted pathways and molecules, model systems continue to increase in importance. The ease of use and versatility of mouse models in the study of human cancer, infectious disease and autoimmunity provides understanding into the etiology of these diseases. Data obtained from studies using mouse models can provide insight into potential clinical side effects and the pathology of diseases.
PBL Assay Science provides a comprehensive mouse product line manufactured to the highest standards of quality, consistency and purity. PBL’s products are designed to meet your scientific needs by providing superior quality and performance to ensure reliable and reproducible results.
Figure 1. Comparison of Mu Alpha A and Mu Alpha 4 Antiviral Activity
Figure 2. Recovery of Mu Alpha A in 50% Serum in Mu Alpha ELISA (42120)
|12100-1||Mouse IFN Alpha A||1 x 105 U|
|12105-1||Mouse IFN Alpha 1||1 x 105 U|
|12115-1||Mouse IFN Alpha 4, mammalian||1 x 105 U|
|12125-1||Mouse IFN Alpha 11||1 x 105 U|
|12130-1||Mouse IFN Alpha 13||1 x 105 U|
|12185-1||NEW C12R Mouse IFN Alpha Decoy Receptor, neutralizing||10 μg|
|12185-2||NEW C12R Mouse IFN Alpha Decoy Receptor, neutralizing||100 μg|
|12400-1||Mouse IFN Beta||1 x 105 U|
|12401-1||Mouse IFN Beta (carrier-free)||1 x 105 U|
|12405-1||Mouse IFN Beta, mammalian||1 x 105 U|
|12410-1||Mouse IFN Beta, mammalian (carrier-free)||1 x 106 U|
|12820-1||Mouse Interleukin-28b/IFN Lambda 3||25 μg|
|12821-1||Mouse Interleukin-28b/IFN Lambda 3 (carrier-free)||25 μg|
|22100-1||Anti-Mouse IFN Alpha, Clone RMMA-1 (MAb)||250 μg|
|22100-3||FITC Conjugated Anti-Mouse IFN Alpha, Clone RMMA-1 (MAb)||25 μg|
|22400-1||Anti-Mouse IFN Beta, Clone RMMB-1 (MAb)||250 μg|
|22400-3||FITC Conjugated Anti-Mouse IFN Beta, Clone RMMB-1 (MAb)||25 μg|
|22500-1||Anti-Mouse IFN Gamma, Clone RMMG-1 (MAb)||500 μg|
|32100-1||Anti-Mouse IFN Alpha, Rabbit Serum (PAb)||2 x 104 U|
|32120-1||Anti-Mouse IFN Alpha, Chicken IgY, Purified (PAb)||500 μg|
|32400-1||Anti-Mouse IFN Beta, Rabbit Serum (PAb)||2 x 104 U|
|32401-1||Anti-Mouse IFN Beta, Rabbit IgG, Protein A Purified (PAb)||2 x 104 U|
|32500-1||Anti-Mouse IFN Gamma, Rabbit Serum (PAb)||2 x 104 U|
|42120-1||VeriKine Mouse IFN Alpha ELISA Kit||1 plate|
|42120-2||VeriKine Mouse IFN Alpha ELISA Kit||5 plate|
|42115-1||NEW VeriKine-HS Mouse IFN Alpha All-Subtype ELISA Kit||1 plate|
|42400-1||VeriKine Mouse IFN Beta ELISA Kit||1 plate|
|42400-2||VeriKine Mouse IFN Beta ELISA Kit||5 plate|
|42410-1||VeriKine-HS Mouse IFN Beta Serum ELISA Kit||1 plate|
|42410-2||VeriKine-HS Mouse IFN Beta Serum ELISA Kit||5 plate|
|52500-1||NEW VeriPlex Mouse Cytokine 9-Plex ELISA Kit||1 plate|
|62830-1||DIY Mouse IFN Lambda 2/3 (IL-28A/B) ELISA||Materials for 15 x 96 wells|
High quality Interferon Gamma products from PBL Assay Science:
Figure 1. Chromatographic profile of IFN-γ showing dimerization and low aggregation
Figure 2. PBL IFN-γ and international gamma standard exhibit comparable antiviral activities
|11500-1||Human IFN Gamma||50 μg|
|11500-2||Human IFN Gamma||100 μg|
|13500-1||Rat IFN Gamma||10 μg|
|21500-1||Anti-Human IFN Gamma, Clone MMHG-1 (MAb)||500 μg|
|21550-1||Anti-Human IFN Gamma, Clone MMHG-2 (MAb)||100 μg|
|21585-1||Anti-Human IFN Gamma Receptor Chain 2, Clone MMHGR-2 (MAb)||250 μg|
|22500-1||Anti-Mouse IFN Gamma, Clone RMMG-1 (MAb)||500 μg|
|31500-1||Anti-Human IFN Gamma, Rabbit Serum (PAb)||2 x 104 U|
|32500-1||Anti-Mouse IFN Gamma, Rabbit Serum (PAb)||2 x 104 U|
|41500-1||VeriKine Human IFN Gamma ELISA Kit||1 plate|
|41580-1||NEW VeriKine Human IFN Gamma Receptor 1 ELISA Kit||1 plate|
Non-human primates are considered a preferred model for the study of human diseases due to a close phylogenetic relationship to humans. The model is crucial to the elucidation of pathological mechanisms and may enable the development of novel diagnostic and therapeutic tools. Studies of innate and adaptive immunity, pharmacodynamics (PD), and immunotoxicology of interferon stimulating/inhibiting molecules, as well as Emerging Infectious Disease (EID) models, can all be further advanced with an interferon immunoassay that is designed specifically for NHP.
PBL now offers ELISAs for the specific detection of NHP IFN-Alpha and IFN-Beta in a variety of sample matrices. Visit our website or contact us for more information on these kits!
VeriKineTM Cynomolgus/Rhesus Interferon Alpha Serum ELISA (Cat. No. 46100-1)
Compatibility: Serum, Plasma, Cell Culture Supernatant
Assay Range: 25 – 1600 pg/ml
Figure 1. Standard Curve in Different Sample Matrices
Figure 2. Specifically designed for detecting NHP IFN-α Greater NHP IFN-α detection sensitivity and matrix compatibility
VeriKineTM Cynomolgus Interferon Beta ELISA (Cat. No. 46415-1)
Compatibility: Serum, Plasma, Cell Culture Supernatant
Assay Range: 5.47 – 350 pg/ml
Figure 3. Typical standard curve from 5.47 to 350 pg/ml
Figure 4. Performance specifications at three different concentrations
Figure 1. Mouse MAb to Human IFN α/β Receptor 2 (MMHAR-2)
Figure 2. Neutralizing MAb against IFNLR1
|10 ng/ml||L1 31.3|
|10 ng/ml||L2 54.3|
|3 ng/ml||L3 22.0|
Figure 3. C12R Neutralization of Mouse IFN α Subtypes on L929 Cells
Figure 4. Neutralization of Human IFN α subtypes on A549 cells
< 1 EU/μg
Carrier protein (PBS with 0.1% BSA) was added to provide extra stability to the cytokine proteins. However, in certain applications, such as in vivo injection, conjugation, or surface binding, carrier protein may interfere.
The formula to use to convert S.A. to pg/ml is as follow: [(1 x 10E+09)/(Lot specific activity in units/mg)] x [Lot-specific concentration in units/ml]
Culture medium supplemented with serum. Interferon beta proteins are unstable at diluted concentrations. Unless formulation work has been done to verify stability, do not dilute carrier-free solution to protein concentration below 100 μg/ml.
Acetic acid prevents aggregation of interferon beta proteins in this particular product formulation.
PBL does not offer any controls for its antibodies at the current time.
For monoclonal antibodies, use normal antibody from the same species and the same isotype. For example, if a MAb is a murine IgG2A, use the normal equivalent as the negative control. Some polyclonal antibodies from PBL are unpurified sera. Therefore, investigators should use normal unpurified serum from the appropriate species as their negative control.
User cell lines vary so there is no specific answer to this question. However, the general rule is 70 fold excess in mass should be used for neutralizing interferon. It is recommended to quantify the amount of interferon via technique such as ELISA. It is also recommended to allow time (1-2 hours in general) for the antibody to neutralize the interferon before adding the virus.
he viral challenge assay is used to test antibodies for neutralization activity. The assay measures the ability of the antibody to negate the protective effects of the interferon against the protective effects of the interferon against the viral challenge.
All PBL ELISA kits are constructed with carefully selected proprietary antibodies and optimized reagents.
PBL offers a range of assay kits that are specifically designed for serum/plasma human interferon alpha (41110, 41115), human interferon beta (41415), mouse interferon beta (42410, 42400), cynomolgus interferon alpha (46100), human IFNs and cytokines (51500).
Due to lot-specific optimization of our ELISA kit reagents, they are not sold separately.
To optimize each lot of kits, PBL modifies the concentration of the reagents to achieve a standard curve that will meet our specific range and signal to noise requirements.
No. Reagent concentration is optimized according to lot. Reagents from different lot can not be used as substitutes without affecting the overall performance of the kit.
Stratech Scientific have been successful in a recent antibodies tender from the Southern Universities Purchasing Consortium. In the award criterion Read More…