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This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human CR1 . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human CR1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human CR1, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human CR1. The concentration of Human CR1 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| Assay type | Sandwich |
| Format | 96T |
| Assay time | 4.5h |
| Reactivity | Human |
| Detection method | Chemiluminescence |
| Detection range | 7.81-500 pg/mL |
| Sensitivity | 4.69 pg/mL |
| Sample volume | 100uL |
| Sample type | Serum, plasma and other biological fluids |
| Repeatability | CV < 15% |
This kit recognizes Human CR1in samples. No significant cross-reactivity or interference between Human CR1and analogues was observed.
As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration(pg/mL) | RLU | Average | Corrected |
|---|---|---|---|
| 500 | 4969953375 | 51537 | 51508 |
| 250 | 1943022566 | 20998 | 20969 |
| 125 | 94849308 | 9396 | 9367 |
| 62.5 | 43804644 | 4512 | 4483 |
| 31.25 | 24352163 | 2299 | 2270 |
| 15.63 | 13271173 | 1250 | 1221 |
| 7.81 | 700780 | 740 | 711 |
| 0 | 2929 | 29 | -- |
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CR1were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CR1were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 25.53 | 43.66 | 198.87 | 26.76 | 47.90 | 211.98 |
| Standard deviation | 2.75 | 4.76 | 14.18 | 3.13 | 3.43 | 21.03 |
| C V (%) | 10.77 | 10.90 | 7.13 | 11.70 | 7.16 | 9.92 |
The recovery of Human CR1spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum (n=5) | 96-111 | 101 |
| EDTA plasma (n=5) | 86-99 | 93 |
| Cell culture media (n=5) | 92-103 | 98 |
Samples were spiked with high concentrations of Human CR1and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 97-109 | 101-115 | 88-104 |
| Average (%) | 103 | 107 | 95 | |
| 1:4 | Range (%) | 99-117 | 91-106 | 98-114 |
| Average (%) | 107 | 97 | 105 | |
| 1:8 | Range (%) | 100-114 | 89-102 | 96-111 |
| Average (%) | 108 | 96 | 103 | |
| 1:16 | Range (%) | 98-115 | 100-117 | 95-110 |
| Average (%) | 106 | 107 | 100 |
An unopened kit can be stored at 4"C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
|---|---|---|
| Micro CLIA Plate(Dismountable) | 8 wells X12 strips | -20"C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100X) | 1 vial, 120 uL | |
| Concentrated HRP Conjugate (100X) | 1 vial, 120 uL | -20"C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4"C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25X) | 1 vial, 30 mL | |
| Substrate Reagent A | 1 vial, 5 mL | 4"C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4"C (shading light) |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
| UniProt Protein Function: | CR1: Mediates cellular binding of particles and immune complexes that have activated complement. Belongs to the receptors of complement activation (RCA) family. |
| UniProt Protein Details: | Protein type:Membrane protein, integral; Receptor, misc. Chromosomal Location of Human Ortholog: 1q32 Cellular Component: cell surface; integral to plasma membrane; plasma membrane Molecular Function:complement component C3b receptor activity; complement component C4b binding; complement component C4b receptor activity; complement component C3b binding Biological Process: negative regulation of complement activation, alternative pathway; complement receptor mediated signaling pathway; negative regulation of complement activation, classical pathway; regulation of complement activation; innate immune response; complement activation, classical pathway Disease: Knops Blood Group System; Malaria, Susceptibility To |
| NCBI Summary: | This gene is a member of the receptors of complement activation (RCA) family and is located in the "cluster RCA" region of chromosome 1. The gene encodes a monomeric single-pass type I membrane glycoprotein found on erythrocytes, leukocytes, glomerular podocytes, and splenic follicular dendritic cells. The Knops blood group system is a system of antigens located on this protein. The protein mediates cellular binding to particles and immune complexes that have activated complement. Decreases in expression of this protein and/or mutations in its gene have been associated with gallbladder carcinomas, mesangiocapillary glomerulonephritis, systemic lupus erythematosus and sarcoidosis. Mutations in this gene have also been associated with a reduction in Plasmodium falciparum rosetting, conferring protection against severe malaria. Alternate allele-specific splice variants, encoding different isoforms, have been characterized. Additional allele specific isoforms, including a secreted form, have been described but have not been fully characterized. [provided by RefSeq, Jul 2008] |
| UniProt Code: | P17927 |
| NCBI GenInfo Identifier: | 86793036 |
| NCBI Gene ID: | 1378 |
| NCBI Accession: | NP_000564.2 |
| UniProt Secondary Accession: | P17927,Q16744, Q16745, Q5SR43, Q5SR45, Q9UQV2, |
| UniProt Related Accession: | P17927 |
| Molecular Weight: | 223,663 Da |
| NCBI Full Name: | complement receptor type 1 isoform F |
| NCBI Synonym Full Names: | complement component (3b/4b) receptor 1 (Knops blood group) |
| NCBI Official Symbol: | CR1 |
| NCBI Official Synonym Symbols: | KN; C3BR; C4BR; CD35 |
| NCBI Protein Information: | complement receptor type 1 |
| UniProt Protein Name: | Complement receptor type 1 |
| UniProt Synonym Protein Names: | C3b/C4b receptor; CD_antigen: CD35 |
| Protein Family: | Complement receptor |
| UniProt Gene Name: | CR1 |
| UniProt Entry Name: | CR1_HUMAN |
Reactivity: | Human |
ELISA Type: | CLIA |
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