Human Cell Biology ELISA Kits 4
Human GHRH (Growth Hormone Releasing Hormone) CLIA Kit (HUES00667)
- SKU:
- HUES00667
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 4.69pg/mL
- Range:
- 7.81-500pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- GRF, GHRF, Somatoliberin, Somatocrinin
- 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: | 7.81-500 pg/mL |
| Sensitivity: | 4.69 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human GHRH in samples. No significant cross-reactivity or interference between Human GHRH 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 GHRH. 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 GHRH 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 GHRH, 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 GHRH. The concentration of Human GHRH in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | GHRH: GRF is released by the hypothalamus and acts on the adenohypophyse to stimulate the secretion of growth hormone. Belongs to the glucagon family. 2 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:Secreted; Secreted, signal peptide Chromosomal Location of Human Ortholog: 20q11. 2 Cellular Component: extracellular space; extracellular region; terminal button Molecular Function:growth hormone-releasing hormone activity; growth hormone-releasing hormone receptor binding Biological Process: response to food; positive regulation of insulin-like growth factor receptor signaling pathway; positive regulation of growth hormone secretion; growth hormone secretion; cAMP-mediated signaling; positive regulation of circadian sleep/wake cycle, REM sleep; cell-cell signaling; positive regulation of cAMP biosynthetic process; positive regulation of cell proliferation; adenohypophysis development; positive regulation of multicellular organism growth; G-protein signaling, adenylate cyclase activating pathway |
| NCBI Summary: | The protein encoded by this gene belongs to the glucagon family and is a preproprotein that is produced in the hypothalamus. The preproprotein is cleaved to form a 44 aa factor, also called somatocrinin, that acts to stimulate growth hormone release from the pituitary. Variant receptors for somatocrinin have been found in several types of tumors, and antagonists of these receptors can inhibit the growth of the tumors. Defects in this gene are a cause of dwarfism, while hypersecretion of the encoded protein is a cause of gigantism. [provided by RefSeq, Jul 2008] |
| UniProt Code: | P01286 |
| NCBI GenInfo Identifier: | 134521 |
| NCBI Gene ID: | 2691 |
| NCBI Accession: | P01286. 1 |
| UniProt Secondary Accession: | P01286,Q4KN10, Q5JYR1, |
| UniProt Related Accession: | P01286 |
| Molecular Weight: | 108 |
| NCBI Full Name: | Somatoliberin |
| NCBI Synonym Full Names: | growth hormone releasing hormone |
| NCBI Official Symbol: | GHRH |
| NCBI Official Synonym Symbols: | GRF; INN; GHRF |
| NCBI Protein Information: | somatoliberin; sermorelin; somatorelin; somatocrinin; growth hormone-releasing factor; growth hormone-releasing hormone |
| UniProt Protein Name: | Somatoliberin |
| UniProt Synonym Protein Names: | Growth hormone-releasing factor; GRF; Growth hormone-releasing hormone; GHRH; Somatocrinin; Somatorelin; INN: Sermorelin |
| Protein Family: | Somatoliberin |
| UniProt Gene Name: | GHRH |
| UniProt Entry Name: | SLIB_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 |
| 500 | 50948 52126 | 51537 | 51508 |
| 250 | 20946 21050 | 20998 | 20969 |
| 125 | 9639 9153 | 9396 | 9367 |
| 62.5 | 4489 4535 | 4512 | 4483 |
| 31.25 | 2337 2261 | 2299 | 2270 |
| 15.63 | 1300 1200 | 1250 | 1221 |
| 7.81 | 700 780 | 740 | 711 |
| 0 | 29 29 | 29 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human GHRH were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human GHRH 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) | 23.57 | 77.03 | 203.32 | 22.06 | 78.33 | 223.46 |
| Standard deviation | 2.19 | 6.59 | 22.20 | 2.61 | 9.25 | 24.85 |
| C V (%) | 9.29 | 8.56 | 10.92 | 11.83 | 11.81 | 11.12 |
Recovery
The recovery of Human GHRH 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) | 93-108 | 99 |
| EDTA plasma (n=5) | 89-101 | 95 |
| Cell culture media (n=5) | 96-114 | 104 |
Linearity
Samples were spiked with high concentrations of Human GHRH 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 (%) | 102-116 | 96-108 | 89-102 |
| Average (%) | 108 | 102 | 94 | |
| 1:4 | Range (%) | 91-107 | 88-100 | 93-106 |
| Average (%) | 98 | 93 | 100 | |
| 1:8 | Range (%) | 95-109 | 89-100 | 96-111 |
| Average (%) | 101 | 94 | 103 | |
| 1:16 | Range (%) | 100-112 | 103-117 | 102-115 |
| Average (%) | 105 | 109 | 108 |
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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