Human Interferon beta / IFN beta ELISA Kit
- SKU:
- HUFI00357
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P01574
- Sensitivity:
- 37.5pg/ml
- Range:
- 62.5-4000pg/ml
- ELISA Type:
- Sandwich
- Synonyms:
- IFN-Beta, IFNB1, Fibroblast interferon, IFBIFNB, IFF, IFN-beta, interferon beta
- Reactivity:
- Human
- Research Area:
- Immunology
Description
Human Interferon beta / IFN beta ELISA Kit
Interferon beta / IFN beta encodes a signaling protein which is released as part of the innate immune response to pathogens. Interferon beta / IFN beta belongs to the type I class of interferons, which are important for defense against viral infections. In addition, Interferon beta / IFN beta is involved in cell differentiation and anti-tumor defenses. Overactivation of Interferon beta / IFN beta is linked to defects in B cell maturation and leads to autoimmune diseases.
Key Features
| Save Time | Pre-coated 96 well plate | |
| Quick Start | Kit includes all necessary reagents | |
| Publication Ready | Reproducible and reliable results |
Overview
| Product Name: | Human Interferon beta / IFN beta ELISA Kit |
| Product Code: | HUFI00357 |
| Size: | 96 Assays |
| Alias: | IFN-beta, IFNB1, Fibroblast interferon, IFBIFNB, IFF, IFN-beta, interferon beta |
| Detection Method: | Sandwich ELISA, Double Antibody |
| Application: | This immunoassay kit allows for the in vitro quantitative determination of Human IFN-beta concentrations in serum plasma and other biological fluids. |
| Sensitivity: | 37.5pg/ml |
| Range: | 62.5-4000pg/ml |
| Storage: | 4°C for 6 months |
| Note: | For Research Use Only |
Additional Information
| Recovery | Matrices listed below were spiked with certain level of Human IFN-beta and the recovery rates were calculated by comparing the measured value to the expected amount of Human IFN-beta in samples.
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| Linearity | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human IFN-beta and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
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| CV(%) | Intra-Assay: CV<8% |
Kit Components
| Component | Quantity | Storage |
| ELISA Microplate (Dismountable) | 8x12 strips | 4°C for 6 months |
| Lyophilized Standard | 2 | 4°C/ -20°C |
| Sample/Standard Dlution Buffer | 20ml | 4°C |
| Biotin-labeled Antibody (Concentrated) | 120ul | 4°C (Protection from light) |
| Antibody Dilution Buffer | 10ml | 4°C |
| HRP-Streptavidin Conjugate (SABC) | 120ul | 4°C (Protect from light) |
| SABC Dilution Buffer | 10ml | 4°C |
| TMB Substrate | 10ml | 4°C (Protection from light) |
| Stop Solution | 10ml | 4°C |
| Wash Buffer (25X) | 30ml | 4°C |
| Plate Sealer | 5 | - |
Other materials required:
- Microplate reader with 450 nm wavelength filter
- Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
- Incubator
- Deionized or distilled water
- Absorbent paper
- Buffer resevoir
Protein Information
Protocol
*Note: Protocols are specific to each batch/lot. For the exact instructions please follow the protocol included in your kit.
Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.
| Step | Procedure |
| 1. | Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells! |
| 2. | Aliquot 0.1ml standard solutions into the standard wells. |
| 3. | Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well. |
| 4. | Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells. |
| 5. | Seal the plate with a cover and incubate at 37 °C for 90 min. |
| 6. | Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2. |
| 7. | Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall. |
| 8. | Seal the plate with a cover and incubate at 37°C for 60 min. |
| 9. | Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash. |
| 10. | Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min. |
| 11. | Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min. |
| 12. | Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color. |
| 13. | Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately. |
| 14. | Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution. |
Sample Type
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
| Sample Type | Protocol |
| Serum | If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. |
| Plasma | Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit. |
| Urine & Cerebrospinal Fluid | Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid. |
| Cell culture supernatant | Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately. |
| Cell lysates | Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Tissue homogenates | The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C. |
| Tissue lysates | Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C |
| Breast Milk | Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles. |
Interferon Beta Background
Interferon Beta
Interferons (IFNs) are a family of naturally occurring proteins that are produced by various cells in response to pathogens, foreign substances, and cancer cells. They were initially discovered six decades ago as antiviral agents synthesized by virus-infected cells. The secretion of IFNs is triggered by a wide range of stimuli and involves cells such as fibroblasts, natural killer cells, white blood cells, and epithelial cells. These proteins play crucial roles in both innate and adaptive immunity, orchestrating immune responses against infections and diseases. Originally categorized into three types (IFNα, IFNβ, and IFNγ), further research has led to the identification of additional forms and their receptors, resulting in the classification of Type I, Type II, and Type III IFNs in humans. Type I IFNs include IFN-α, IFN-β, IFN-É›, IFN-κ, and IFN-ω, while Type II consists of IFN-γ, and Type III comprises the IFNλ subtypes.
Interferon Beta Structure
Interferon beta exhibits a unique structural composition that contributes to its diverse functions in the immune system. It belongs to the type I interferon family, characterized by its three-dimensional arrangement of amino acids. Interferon beta consists of a single polypeptide chain containing 166 amino acids, organized into five distinct structural domains. The N-terminal domain forms a signal peptide essential for proper secretion of Interferon beta. This is followed by a conserved cysteine-rich domain involved in protein stability and receptor binding. The central alpha-helical domain facilitates protein-protein interactions, while the C-terminal domain plays a role in receptor engagement and signaling. Lastly, the glycosylation sites within the protein contribute to its stability and bioactivity. Understanding the intricate structure of Interferon beta is crucial for elucidating its mechanisms of action and designing targeted therapies to harness its therapeutic potential.
Role of IFN-Beta in Treatment of Diseases
The therapeutic potential of Interferon beta has been extensively studied and harnessed for the treatment of various diseases. Its remarkable ability to modulate immune responses and exhibit antiviral properties has made it a valuable tool in combating conditions such as multiple sclerosis (MS). In the context of MS, Interferon beta helps regulate the immune system by reducing the inflammatory response and inhibiting the activation of autoreactive T cells, thereby slowing down the progression of the disease and reducing relapse rates. Additionally, Interferon beta has shown promise in the treatment of viral infections, including hepatitis B and C, by boosting the body's natural defense mechanisms against these viruses. Furthermore, its immunomodulatory effects have been explored in the treatment of certain types of cancers, where it aids in stimulating antitumor responses. The multifaceted role of Interferon beta in disease management highlights its potential as a therapeutic agent in various medical conditions, paving the way for further research and advancements in treatment strategies.
Human IFN-Beta ELISA Kit FAQs
What is the Human IFN-Beta ELISA Kit used for?
The Human IFN-Beta ELISA Kit is a specialized tool used for the quantitative measurement of Interferon Beta (IFN-Beta) levels in various biological samples. It is commonly employed in immunological research and diagnostics to gain insights into allergic reactions and related diseases.
What are the advantages of using the IFN-Beta ELISA Kit?
The Human IFN-Beta ELISA Kit offers several advantages, including high sensitivity, accuracy, and reproducibility. It provides a user-friendly and reliable method to quantify IFN-Beta levels in biological specimens, allowing for precise measurements and robust data analysis.
What sample types are compatible with the IFN-Beta ELISA Kit?
The IFN-Beta ELISA Kit is compatible with various sample types, including serum, plasma, cell lysates, and tissue homogenates. It provides flexibility in sample selection, allowing researchers to analyze IFN-Beta levels in different biological matrices.
What are the storage requirements for the IFN-Bet ELISA Kit?
The IFN-Beta ELISA Kit components should be stored according to the instructions provided in the kit manual. Generally, it is recommended to store the kit components at the recommended temperature to ensure their stability and optimal performance.
What should I do if my assay results are not optimal?
If you encounter any issues or have suboptimal assay results, we recommend contacting our dedicated support team for assistance. They will be available to provide troubleshooting guidance, answer your questions, and ensure you achieve the best possible results with the Fc epsilon RI ELISA Kit.
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