Description
Human Complement factor D (CFD) ELISA Kit
The Human Complement Factor D (CFD) ELISA Kit is specifically designed for the precise and reliable quantification of complement factor D levels in human samples including serum, plasma, and cell culture supernatants. This ELISA kit offers high sensitivity and specificity, ensuring accurate and reproducible results for a wide range of research applications.Complement factor D is an important component of the alternative pathway of the complement system, playing a crucial role in the immune response against pathogens and contributing to inflammatory processes. Abnormal levels of complement factor D have been implicated in various diseases, including autoimmune disorders, infectious diseases, and inflammatory conditions, making it a valuable biomarker for disease diagnosis and therapeutic development.
With the Human Complement Factor D (CFD) ELISA Kit, researchers can effectively study the role of complement factor D in health and disease, advancing our understanding of immune responses and potential therapeutic interventions. Order your kit today and enhance your research efforts with reliable and accurate quantification of complement factor D levels.
| Product Name: | Human Complement factor D (CFD) ELISA Kit |
| SKU: | HUEB0289 |
| Size: | 96T |
| Target: | Human Complement factor D (CFD) |
| Synonyms: | Adipsin, C3 convertase activator, Properdin factor D, DF, PFD |
| Assay Type: | Sandwich |
| Detection Method: | ELISA |
| Reactivity: | Human |
| Detection Range: | 7.8-500pg/mL |
| Sensitivity: | 3.9 pg/mL |
| Intra CV: | 3.8% | ||||||||||||||||||||
| Inter CV: | 7.5% | ||||||||||||||||||||
| Linearity: |
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| Recovery: |
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| Function: | Factor D cleaves factor B when the latter is complexed with factor C3b, activating the C3bbb complex, which then becomes the C3 convertase of the alternate pathway. Its function is homologous to that of C1s in the classical pathway. |
| Uniprot: | P00746 |
| Sample Type: | Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids |
| Specificity: | Natural and recombinant human Complement factor D |
| Research Area: | Immunology |
| Subcellular Location: | Secreted |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| UniProt Protein Function: | CFD: Factor D cleaves factor B when the latter is complexed with factor C3b, activating the C3bbb complex, which then becomes the C3 convertase of the alternate pathway. Its function is homologous to that of C1s in the classical pathway. Defects in CFD are the cause of complement factor D deficiency (CFDD). CFDD is an immunologic disorder characterized by increased susceptibility to bacterial infections, particularly Neisseria infections, due to a defect in the alternative complement pathway. Belongs to the peptidase S1 family. |
| UniProt Protein Details: | Protein type:Secreted; EC 3.4.21.46; Secreted, signal peptide; Protease Chromosomal Location of Human Ortholog: 19p13.3 Cellular Component: extracellular region Molecular Function:serine-type endopeptidase activity; serine-type peptidase activity Biological Process: complement activation; complement activation, alternative pathway; platelet degranulation Disease: Complement Factor D Deficiency |
| NCBI Summary: | This gene encodes a member of the S1, or chymotrypsin, family of serine peptidases. This protease catalyzes the cleavage of factor B, the rate-limiting step of the alternative pathway of complement activation. This protein also functions as an adipokine, a cell signaling protein secreted by adipocytes, which regulates insulin secretion in mice. Mutations in this gene underlie complement factor D deficiency, which is associated with recurrent bacterial meningitis infections in human patients. Alternative splicing of this gene results in multiple transcript variants. At least one of these variants encodes a preproprotein that is proteolytically processed to generate the mature protease. [provided by RefSeq, Nov 2015] |
| UniProt Code: | P00746 |
| NCBI GenInfo Identifier: | 158515408 |
| NCBI Gene ID: | 1675 |
| NCBI Accession: | P00746.5 |
| UniProt Secondary Accession: | P00746,Q5U5S1, Q86VJ5, Q8N4E0, Q8WZB4, B4DV76, |
| UniProt Related Accession: | P00746 |
| Molecular Weight: | 27,033 Da |
| NCBI Full Name: | Complement factor D |
| NCBI Synonym Full Names: | complement factor D |
| NCBI Official Symbol: | CFD |
| NCBI Official Synonym Symbols: | DF; ADN; PFD; ADIPSIN |
| NCBI Protein Information: | complement factor D |
| UniProt Protein Name: | Complement factor D |
| UniProt Synonym Protein Names: | Adipsin; C3 convertase activator; Properdin factor D |
| Protein Family: | Complement factor |
| UniProt Gene Name: | CFD |
| UniProt Entry Name: | CFAD_HUMAN |
| Component | Quantity (96 Assays) | Storage |
| ELISA Microplate (Dismountable) | 8×12 strips | -20°C |
| Lyophilized Standard | 2 | -20°C |
| Sample Diluent | 20ml | -20°C |
| Assay Diluent A | 10mL | -20°C |
| Assay Diluent B | 10mL | -20°C |
| Detection Reagent A | 120µL | -20°C |
| Detection Reagent B | 120µL | -20°C |
| Wash Buffer | 30mL | 4°C |
| Substrate | 10mL | 4°C |
| Stop Solution | 10mL | 4°C |
| Plate Sealer | 5 | - |
Other materials and equipment 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
*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.
| Step | |
| 1. | Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C. |
| 2. | Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform. |
| 3. | Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper. |
| 4. | Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C. |
| 5. | Repeat the wash process for five times as conducted in step 3. |
| 6. | Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction. |
| 7. | Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing. |
| 8. | Determine the optical density (OD value) of each well at once, using a micro-plate reader |