Human PTHrP (Parathyroid Hormone Related Protein) ELISA Kit (HUES02506)
- Product Type:
- ELISA Kit
- 96 Assays
- ELISA Type:
- Tested Sample Types:
- Serum, plasma and other biological fluids
|Detection Range:||15.63-1000 pg/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Human PTHrP in samples. No significant cross-reactivity or interference between Human PTHrP and analogues was observed.|
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human PTHrP. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human PTHrP 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 PTHrP, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding 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 Human PTHrP. The concentration of Human PTHrP in samples can be calculated by comparing the OD of the samples to the standard curve.
|UniProt Protein Function:||PTHrP: a member of the parathyroid hormone family. This hormone regulates endochondral bone development and epithelial-mesenchymal interactions during the formation of the mammary glands and teeth. This hormone is involved in lactation possibly by regulating the mobilization and transfer of calcium to the milk. The receptor of this hormone, PTHR1, is responsible for most cases of humoral hypercalcemia of malignancy. Three alternatively spliced isoforms have been described.|
|UniProt Protein Details:|
Protein type:Secreted; Secreted, signal peptide
Chromosomal Location of Human Ortholog: 12p12. 1-p11. 2
Cellular Component: nucleoplasm; Golgi apparatus; extracellular space; cytoplasm; extracellular region
Molecular Function:peptide hormone receptor binding; hormone activity
Biological Process: epidermis development; negative regulation of transcription factor activity; negative regulation of chondrocyte differentiation; female pregnancy; G-protein signaling, adenylate cyclase activating pathway; osteoblast development; negative regulation of cell proliferation; cAMP metabolic process; epithelial cell differentiation; cell-cell signaling; regulation of gene expression; positive regulation of cell proliferation; positive regulation of cAMP biosynthetic process; protein processing; skeletal development; surfactant homeostasis; alveolus development; endochondral ossification; endoderm development
Disease: Brachydactyly, Type E2
|NCBI Summary:||The protein encoded by this gene is a member of the parathyroid hormone family. This hormone, via its receptor, PTHR1, regulates endochondral bone development and epithelial-mesenchymal interactions during the formation of the mammary glands and teeth. It is responsible for most cases of humoral hypercalcemia of malignancy, and mutations in this gene are associated with brachydactyly type E2 (BDE2). Alternatively spliced transcript variants have been found for this gene. There is also evidence for alternative translation initiation from non-AUG (CUG and GUG) start sites, downstream of the initiator AUG codon, resulting in nuclear forms of this hormone. [provided by RefSeq, Nov 2013]|
|NCBI GenInfo Identifier:||131542|
|NCBI Gene ID:||5744|
|NCBI Accession:||P12272. 1|
|UniProt Secondary Accession:||P12272,Q15251, Q6FH74,|
|UniProt Related Accession:||P12272|
|Molecular Weight:||Calculated MW: 19kDa/20kDa/23kDa|
|NCBI Full Name:||Parathyroid hormone-related protein|
|NCBI Synonym Full Names:||parathyroid hormone-like hormone|
|NCBI Official Symbol:||PTHLH|
|NCBI Official Synonym Symbols:||HHM; PLP; BDE2; PTHR; PTHRP|
|NCBI Protein Information:||parathyroid hormone-related protein; PTH-rP; osteostatin; PTH-related protein; parathyroid hormone-like related protein; parathyroid hormone-like hormone preproprotein|
|UniProt Protein Name:||Parathyroid hormone-related protein|
|UniProt Synonym Protein Names:||Parathyroid hormone-like protein; PLP|
|Protein Family:||Parathyroid hormone-related protein|
|UniProt Gene Name:||PTHLH|
|UniProt Entry Name:||PTHR_HUMAN|
As the OD 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.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PTHrP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PTHrP were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||6.72||4.24||4.89||5.03||5.03||5.06|
The recovery of Human PTHrP spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||94-106||99|
|Cell culture media (n=5)||87-101||94|
Samples were spiked with high concentrations of Human PTHrP 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)|
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.
|Micro ELISA 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||1 vial, 10 mL||4°C(shading light)|
|Stop Solution||1 vial, 10 mL||4°C|
|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 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.