KEY FEATURES
| Full Name | Complement Component 4a |
|---|---|
| Synonym | C4a anaphylatoxin |
| Assay Type | Sandwich |
| Reactivity | Mouse |
| Range | 31.2-2000pg/mL |
| Sensitivity | 13.3pg/mL |
| Sample Type | Serum,plasma and other biological fluids |
| Sample Volume | 100μL |
| Detection Wavelength | OD450 |
| Transportation Temperature | 2-8°C |
| Specificity | The kit detected Mouse C4a in the samples and no significant cross-species reactions were found |
| Microplate | 96-wells plate breakable into 12 x 8 wells strip |
TEST PRINCIPLE
| This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse C4a. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse C4a and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse C4a, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse C4a. You can calculate the concentration of Mouse C4a in the samples by comparing the OD of the samples to the standard curve. |
ELISA KIT COMPONENTS
Upon receipt, unpack promptly and store as recommended in the instructions.
| Components | Specifications | Storage and Notes |
|---|---|---|
| Micro Plate | 96T: 8 wells×12 strips 48T: 8 wells×6 strips | Unopened: -20°C, 12 months Unused: Put it back in the aluminum foil bag and seal it, store it at -20°C. |
| Reference Standard | 96T: 2 vials 48T: 1 vial | Unopened: -20°C, 12 months Please use freshly dissolved standards for each experiment. Discard any unused standards after dissolution. |
| Biotinylated Detection Ab Concentrate (100×) | 96T: 120μL×1 vial 48T: 60μL×1 vial | Unopened: -20°C, 12 months Unused: Please seal the concentrate and store it at -20°C, and discard the working solution. |
| HRP Conjugate Concentrate (100×) | 96T: 120μL×1 vial 48T: 60μL×1 vial | Unopened: -20°C(Protect from light), 12 months Unused: Please seal the concentrate and store it at -20°C, and discard the working solution. |
| Biotinylated Detection Ab Diluent | 14mL×1 | 2-8℃, 12 months |
| HRP Conjugate Diluent | 14mL×1 | 2-8℃, 12 months |
| Reference Standard & Sample Diluent | 20mL×1 | 2-8℃, 12 months |
| Washing Buffer Concentrate (25×) | 30mL×1 | 2-8℃, 12 months |
| Substrate Reagent(TMB) | 10mL×1 | 2-8°C(Protect from light),12 months |
| Stop Solution | 7mL×1 | 2-8°C/Room temperature |
ASSAY PROCEDURES
TYPICAL DATA
Mouse C4a ELISA Standard Curve
Typical data are for reference only and curves should be replotted for each experiment. The Logistics function is recommended for fitting.
PRECISION
Intra-Assay Precision (Precision within an assay): Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays): Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 105.7 | 269.54 | 742.07 | 108.49 | 268.71 | 745.78 |
| Standard deviation | 5.42 | 13.5 | 32.65 | 6.82 | 15.53 | 37.96 |
| CV(%) | 5.13 | 5.01 | 4.4 | 6.29 | 5.78 | 5.09 |
RECOVERY
The recovery of Mouse C4a spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum(n=8) | 87-99 | 94 |
| EDTA plasma (n=8) | 95-111 | 102 |
| Cell culture media (n=8) | 91-105 | 98 |
LINEARITY
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Mouse C4a in various matrices were diluted with the Reference Standard & Sample Diluent to produce samples with values within the dynamic range of the assay.
Products associated with C4a ELISA Kits
| C1S ELISA Kit | publications with C4a and C1S |
| TNXA ELISA Kit | publications with C4a and TNXA |
| CYP21A2 ELISA Kit | publications with C4a and CYP21A2 |
| C8A ELISA Kit | publications with C4a and C8A |
| HLA-ABC ELISA Kit | publications with C4a and HLA-ABC |
| C5AR1 ELISA Kit | publications with C4a and C5AR1 |
Pathways associated with C4a ELISA Kit
| Activation Of C3 And C5 Pathway | Activation Of C3 And C5 Pathway |
| Allograft Rejection Pathway | Allograft Rejection Pathway |
| Complement Activation, Classical Pathway | Complement Activation, Classical Pathway |
| Complement And Coagulation Cascades Pathway | Complement And Coagulation Cascades Pathway |
| Complement Cascade Pathway | Complement Cascade Pathway |
| Immune System Pathway | Immune System Pathway |
| Initial Triggering Of Complement Pathway | Initial Triggering Of Complement Pathway |
| Innate Immune System Pathway | Innate Immune System Pathway |
| Pertussis Pathway | Pertussis Pathway |
Diseases associated with C4a ELISA Kit
| Lupus Erythematosus, Systemic | publications with C4a and Lupus Erythematosus, Systemic |
| Inflammation | publications with C4a and Inflammation |
| Kidney Diseases | publications with C4a and Kidney Diseases |
| Hyperplasia | publications with C4a and Hyperplasia |
| Disorders of Sex Development | publications with C4a and Disorders of Sex Development |
| Urogenital Abnormalities | publications with C4a and Urogenital Abnormalities |
| Liver Diseases | publications with C4a and Liver Diseases |
| Necrosis | publications with C4a and Necrosis |
| Heart Diseases | publications with C4a and Heart Diseases |
| Hepatitis | publications with C4a and Hepatitis |
Organs/Tissues associated with C4a ELISA Kit
| Blood | publications with C4a and Blood |
| Liver | publications with C4a and Liver |
| Kidney | publications with C4a and Kidney |
| Mouth | publications with C4a and Mouth |
| Heart | publications with C4a and Heart |
| Lung | publications with C4a and Lung |
| Eye | publications with C4a and Eye |
| Muscle | publications with C4a and Muscle |
| Brain | publications with C4a and Brain |
| Adrenal Gland | publications with C4a and Adrenal Gland |
