Human FcER1 / Fc epsilon RI ELISA Kit (HUFI02067)
- SKU:
- HUFI02067
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P12319
- Sensitivity:
- 18.75pg/ml
- Range:
- 31.25-2000pg/ml
- ELISA Type:
- Sandwich
- Synonyms:
- FCER1A, FCE1A, FcERI, Fc-epsilon RI-alpha, IgE Fc receptor subunit alpha,
- Reactivity:
- Human
- Research Area:
- Cell Biology
Description
Human FcER1 / Fc epsilon RI ELISA Kit
FcER1 / Fc epsilon RI binds to the Fc region of immunoglobulins epsilon, and is responsible for initiating the allergic response. FcER1 / Fc epsilon RI receptor also induces the secretion of important lymphokines. Diseases associated with FcER1 / Fc epsilon RI include chronic spontaneous urticaria. The Assay Genie Human FcER1 / Fc epsilon RI ELISA Kit is a highly sensitive assay for the quantitative measurement of FcER1 / Fc epsilon RI in serum, plasma, cell culture supernatant and tissue samples.
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 FcER1 / Fc epsilon I ELISA Kit |
| Product Code: | HUFI02067 |
| Size: | 96 Assays |
| Alias: | FCER1A, FCE1A, FcER1, Fc-Epsilon RI-alpha, IgE Fc receptor subunit alpha |
| Detection Method: | Sandwich ELISA, Double Antibody |
| Application: | This immunoassay kit allows for the in virto quantitative determination of Human FCER1A concetrations in serum plasma and the other biological fluids. |
| Sensitivity: | 18.75pg/ml |
| Range: | 31.25-2000pg/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 FCER1A and the recovery rates were calculated by comparing the measured value to the expected amount of Human FCER1 in samples.
| Matrix | Recovery Range (%) | Average (%) |
| serum (n=5) | 85-98 | 93 |
| EDTA plasma (n=5) | 86-104 | 99 |
| UFH plasma (n=5) | 89-105 | 96 |
Linearity:
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human FCER1A and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Sample | 1:2 | 1:4 | 1:8 |
| serum (n=5) | 91-105% | 87-105% | 89-104% |
| EDTA plasma (n=5) | 88-100% | 89-99% | 85-99% |
| UFH plasma (n=5) | 87-97% | 81-97% | 81-98% |
CV(%):
Intra - Assay: CV<8%
Intra - Assay: CV<10%
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
| UniProt | |
| UniProt Protein Function: | Binds to Fc region of immunoglobulins epsilon. High affinity receptor. Responsible for initiating the allergic response. Binding of allergen to receptor-bound IgE leads to cell activation and the release of mediators (such as histamine) responsible for the manifestations of allergy. The same receptor also induces the secretion of important lymphokines. |
| NCBI Summary: | The immunoglobulin epsilon receptor (IgE receptor) is the initiator of the allergic response. When two or more high-affinity IgE receptors are brought together by allergen-bound IgE molecules, mediators such as histamine that are responsible for allergy symptoms are released. This receptor is comprised of an alpha subunit, a beta subunit, and two gamma subunits. The protein encoded by this gene represents the alpha subunit. [provided by RefSeq, Aug 2011] |
| UniProt Code: | |
| NCBI GenInfo Identifier: | |
| NCBI Gene ID: | |
| NCBI Accession: | |
| UniProt Related Accession: | |
| Molecular Weight: | 30kDa |
| NCBI Full Name: | High affinity immunoglobulin epsilon receptor subunit alpha |
| NCBI Synonym Full Names: | Fc fragment of IgE, high affinity I, receptor for; alpha polypeptide |
| NCBI Official Symbol: | FCER1A |
| NCBI Official Synonym Symbols: | FCE1A; FcERI |
| NCBI Protein Information: | high affinity immunoglobulin epsilon receptor subunit alpha; Fc-epsilon RI-alpha; Fc epsilon RI alpha-chain; igE Fc receptor subunit alpha; Fc IgE receptor, alpha polypeptide; high affinity immunoglobulin epsilon receptor alpha-subunit; immunoglobulin E receptor, high-affinity, of mast cells, alpha polypeptide |
| UniProt Protein Name: | High affinity immunoglobulin epsilon receptor subunit alpha |
| UniProt Synonym Protein Names: | Fc-epsilon RI-alpha; FcERI; IgE Fc receptor subunit alpha |
| Protein Family: | High affinity immunoglobulin epsilon receptor |
| UniProt Gene Name: | FCER1A |
| UniProt Entry Name: | FCERA_HUMAN |
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 Preparation
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. |
FcER1 Background
FcER1
FcER1, also known as Fc epsilon receptor 1, is a crucial component of the high-affinity receptor for immunoglobulin E (IgE). It is a transmembrane protein that is predominantly expressed on mast cells, basophils, and antigen-presenting cells. FcER1 plays a central role in allergic reactions and immune responses by binding to IgE antibodies, which leads to the activation of downstream signaling pathways and the release of inflammatory mediators. Through its interaction with IgE, FcER1 initiates a cascade of events that contribute to the development of allergic symptoms such as itching, swelling, and bronchoconstriction. Understanding the function and regulation of FcER1 is essential for unraveling the mechanisms underlying allergic diseases and for the development of targeted therapeutic interventions.
FcER1 Signaling Pathway
The Fc epsilon RI signaling pathway is a complex molecular cascade that plays a pivotal role in the immune response, particularly in allergic reactions. Upon binding to allergens, immunoglobulin E (IgE) antibodies attached to Fc epsilon RI receptors on the surface of mast cells and basophils trigger a series of intracellular events. This initiates a signaling cascade involving the activation of key proteins, including Syk kinase, LAT (linker for activation of T cells), and PLC-gamma (phospholipase C-gamma). These proteins act as molecular messengers, transmitting signals to downstream effectors, leading to the release of inflammatory mediators such as histamine, leukotrienes, and cytokines. The Fc epsilon RI signaling pathway plays a crucial role in mediating allergic responses, contributing to the development of symptoms such as itching, swelling, and bronchoconstriction. Understanding the intricacies of this signaling pathway is essential for designing targeted therapies aimed at modulating allergic reactions and developing treatments for allergic diseases.
FcER1A Antibody
The FCER1A antibody is a valuable research tool widely used in immunology studies and diagnostics. FCER1A, also known as the Fc epsilon RI alpha subunit, is a key component of the high-affinity receptor for immunoglobulin E (IgE). This transmembrane protein is primarily expressed on mast cells, basophils, and antigen-presenting cells. The FCER1A antibody specifically targets and recognizes the FCER1A protein, allowing researchers to detect, quantify, and visualize its expression and localization in various biological samples. By utilizing the FCER1A antibody, scientists can gain insights into the regulation and activation of the Fc epsilon RI complex, which is central to allergic reactions and immune responses. The FCER1A antibody is an essential tool for studying the role of FCER1A in disease mechanisms, identifying potential therapeutic targets, and advancing our understanding of allergic diseases and immune disorders.
FcER1 ELISA Kit FAQs
What is the Fc epsilon RI ELISA Kit used for?
The Fc epsilon RI ELISA Kit is used to measure the levels of Fc epsilon RI, the high-affinity receptor for immunoglobulin E (IgE), 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 Fc epsilon RI ELISA Kit?
The Fc epsilon RI ELISA Kit offers several advantages, including high sensitivity, accuracy, and reproducibility. It provides a user-friendly and reliable method to quantify Fc epsilon RI levels in biological specimens, allowing for precise measurements and robust data analysis.
What sample types are compatible with the Fc epsilon RI ELISA Kit?
The Fc epsilon RI 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 Fc epsilon RI levels in different biological matrices.
What are the storage requirements for the Fc epsilon RI ELISA Kit?
The Fc epsilon RI 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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