Human Cell Biology ELISA Kits 4
Human PCDH beta16 (Protocadherin Beta 16) CLIA Kit (HUES00558)
- SKU:
- HUES00558
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 18.75pg/mL
- Range:
- 31.25-2000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- PCDHB16, ME1, PCDH-BETA16, PCDH3X, PCDHB8a
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Biology
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection method: | Chemiluminescence |
| Detection range: | 31.25-2000 pg/mL |
| Sensitivity: | 18.75 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human PCDH beta16 in samples. No significant cross-reactivity or interference between Human PCDH beta16 and analogues was observed. |
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 PCDH beta16. 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 PCDH beta16 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 PCDH beta16, 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 PCDH beta16. The concentration of Human PCDH beta16 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | PCDHB16: Potential calcium-dependent cell-adhesion protein. May be involved in the establishment and maintenance of specific neuronal connections in the brain. |
| UniProt Protein Details: | Chromosomal Location of Human Ortholog: 5q31 Cellular Component: integral to membrane; plasma membrane Molecular Function:calcium ion binding Biological Process: synaptic transmission; synaptogenesis; calcium-dependent cell-cell adhesion; homophilic cell adhesion |
| NCBI Summary: | This gene is a member of the protocadherin beta gene cluster, one of three related gene clusters tandemly linked on chromosome five. The gene clusters demonstrate an unusual genomic organization similar to that of B-cell and T-cell receptor gene clusters. The beta cluster contains 16 genes and 3 pseudogenes, each encoding 6 extracellular cadherin domains and a cytoplasmic tail that deviates from others in the cadherin superfamily. The extracellular domains interact in a homophilic manner to specify differential cell-cell connections. Unlike the alpha and gamma clusters, the transcripts from these genes are made up of only one large exon, not sharing common 3' exons as expected. These neural cadherin-like cell adhesion proteins are integral plasma membrane proteins. Their specific functions are unknown but they most likely play a critical role in the establishment and function of specific cell-cell neural connections. [provided by RefSeq, Jul 2008] |
| UniProt Code: | Q9NRJ7 |
| NCBI GenInfo Identifier: | 605052038 |
| NCBI Gene ID: | 57717 |
| NCBI Accession: | NP_066008. 2 |
| UniProt Secondary Accession: | Q9NRJ7,Q8IYD5, Q96SE9, Q9HCF1, B3KPK5, |
| UniProt Related Accession: | Q9NRJ7 |
| Molecular Weight: | 84,983 Da |
| NCBI Full Name: | protocadherin beta-16 |
| NCBI Synonym Full Names: | protocadherin beta 16 |
| NCBI Official Symbol: | PCDHB16 |
| NCBI Official Synonym Symbols: | ME1; PCDH3X; PCDHB8a; PCDH-BETA16 |
| NCBI Protein Information: | protocadherin beta-16; cadherin ME1; protocadherin beta 8a; protocadherin-3x |
| UniProt Protein Name: | Protocadherin beta-16 |
| UniProt Synonym Protein Names: | Protocadherin-3X |
| Protein Family: | Protocadherin |
| UniProt Gene Name: | PCDHB16 |
| UniProt Entry Name: | PCDBG_HUMAN |
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 |
| 2000 | 54655 56787 | 55721 | 55693 |
| 1000 | 23367 24581 | 23974 | 23946 |
| 500 | 11437 10627 | 11032 | 11004 |
| 250 | 5227 5361 | 5294 | 5266 |
| 125 | 2751 2465 | 2608 | 2580 |
| 62.5 | 1354 1268 | 1311 | 1283 |
| 31.25 | 644 704 | 674 | 646 |
| 0 | 27 29 | 28 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PCDH beta16 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PCDH beta16 were 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) | 103.27 | 303.65 | 718.47 | 96.97 | 291.61 | 709.93 |
| Standard deviation | 10.36 | 35.71 | 51.87 | 11.09 | 22.25 | 63.54 |
| C V (%) | 10.03 | 11.76 | 7.22 | 11.44 | 7.63 | 8.95 |
Recovery
The recovery of Human PCDH beta16 spiked 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) | 91-107 | 98 |
| EDTA plasma (n=5) | 95-109 | 102 |
| Cell culture media (n=5) | 96-111 | 101 |
Linearity
Samples were spiked with high concentrations of Human PCDH beta16 and 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 (%) | 91-105 | 96-109 | 100-116 |
| Average (%) | 98 | 101 | 109 | |
| 1:4 | Range (%) | 100-116 | 101-115 | 96-113 |
| Average (%) | 108 | 109 | 103 | |
| 1:8 | Range (%) | 92-107 | 86-97 | 102-116 |
| Average (%) | 99 | 91 | 108 | |
| 1:16 | Range (%) | 92-108 | 100-114 | 94-108 |
| Average (%) | 98 | 106 | 99 |
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 ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -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 (25×) | 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 |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids. ) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
- Determine the RLU value of each well immediately.
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