Human IL-6 ELISA Kit
- SKU:
- HUFI00180
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P05231
- Sensitivity:
- 2.813pg/ml
- Range:
- 4.688-300pg/ml
- ELISA Type:
- Sandwich
- Synonyms:
- IL-6, Interleukin 6, IL6, BSF2, HGF, HSF, IFNB2, CDF
- Reactivity:
- Human
Description
Human IL-6 ELISA Kit
IL-6 ELISA kits are used to detect Interleukin 6 in biological samples. IL-6 plays an important role in the immune response against infections, as it functions to regulate immune cells such as activated T and B lymphocytes and macrophages, by promoting proliferation and differentiation into their activated form. IL-6 is also involved in the proliferation of B cells and production of immunoglobulins IgM, IgG and IgA by these cells. IL-6 is an important biomarker for immune system activity, which has implications in the development of cancer, autoimmune disease and sepsis. IL-6 concentration may be elevated in patients suffering from immune diseases such as rheumatoid arthritis or systemic lupus erythematosus. It may also be elevated during bacterial and viral infections, as IL-6 helps in the immune response against these pathogens.
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 IL-6 ELISA Kit |
| Product Code: | HUFI00180 |
| Size: | 96 Assays |
| Alias: | IL-6, Interleukin 6, IL6, BSF2, HGF, HSF, IFNB2, CDF |
| Detection Method: | Sandwich ELISA, Double Antibody |
| Application: | Quantitative determination of IL-6 concentrations in serum plasma and other biological fluids. |
| Sensitivity: | 2.813pg/ml |
| Range: | 4.688-300pg/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 IL-6 and the recovery rates were calculated by comparing the measured value to the expected amount of Human IL-6 in samples.
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| Linearity: | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human IL-6 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
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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
| Heading 1 | Heading 2 |
| UniProt Code | |
| Protein Family | Interleukin |
| NCBI Full Name | Interleukin-6 |
| NCBI Official Symbol | IL6 |
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. |
IL-6 Background
Interleukin 6
Interleukin 6 (IL-6) is a protein with a molecular weight of 26 kDa, which is secreted by cells. The mature form of IL-6 consists of 184 amino acids and contains two N-glycosylation sites as well as four cysteine residues. It serves as a multifunctional cytokine, exerting its effects in various biological systems and organs. Upon the detection of pathogens at the site of infection or tissue injury by toll-like receptors (TLRs), myeloid cells such as macrophages and dendritic cells produce IL-6. Initially identified as B-cell stimulatory factor 2 (BSF-2) due to its ability to enhance immunoglobulin synthesis in activated B cells, IL-6 is promptly generated to contribute to host defense mechanisms during infections or tissue injuries. However, excessive production of IL-6 has been associated with the development of severe and life-threatening complications, such as systemic inflammatory response syndrome (SIRS) and cytokine-release syndrome (CRS). Notably, IL-6 is the predominant cytokine involved in the cytokine storm observed in certain patients following chimeric antigen receptor (CAR) T-cell therapy.
Interleukin 6 Function
Interleukin-6 (IL-6) is a versatile cytokine with diverse functions that are vital for the body's defense mechanisms. It exerts a significant impact on immune and hematopoietic processes and possesses a strong capability to trigger the acute phase response. The production of IL-6 has both pro- and anti-inflammatory consequences, underscoring its pivotal role in activating and modulating immune responses. Some of the physiological roles of IL-6 include regulating the differentiation of monocytes into macrophages by influencing the expression of macrophage colony-stimulating factor, enhancing IgG production from activated B cells through the regulation of IL-21 expression, inhibiting the maturation of dendritic cells by activating the STAT3 signaling pathway, and promoting the Th2 response by suppressing Th1 polarization.
Interleukin 6 Receptor
The IL-6 receptor complex is composed of two main components: the IL-6-specific α receptor called IL-6Rα (also known as gp80 or CD126) and the signal transducing subunit known as glycoprotein 130 (also referred to as gp130 or CD130). IL-6 initially attaches to IL-6Rα with relatively weak binding.
IL-6 ELISA Kit FAQs
What is the IL-6 ELISA Kit used for?
The Human IL-6 ELISA Kit is specifically designed for the quantitative measurement of interleukin-6 (IL-6) levels in human biological samples. IL-6 is a cytokine that plays a crucial role in immune response regulation and inflammation. This kit enables researchers and clinicians to accurately assess IL-6 concentrations, facilitating investigations into various diseases and conditions associated with IL-6 dysregulation.
What are the advantages of using the IL-6 ELISA Kit?
The IL-6 ELISA Kit offers several advantages, including high sensitivity, accuracy, and reproducibility. It provides a user-friendly and reliable method to quantify IL-6 levels in biological specimens, allowing for precise measurements and robust data analysis
What sample types are compatible with the IL-6 ELISA Kit?
The IL-6 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 IL-6 levels in different biological matrices.
What are the storage requirements for the IL-6 ELISA Kit?
The IL-6 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 Il-6 ELISA Kit.
Citations
| Morris et al. | Decreased expression of the translation factor eIF3e induces senescence in breast cancer cells via suppression of PARP1 and activation of mTORC1 | Oncotarget. (2021) | PubMed: 33868586 |
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