ELISA Kit for Parathyroid Hormone Related Protein (PTHrP) Mus musculus (Mouse) Competition ELISA

PTHLH; HHM; PTHR; PLP; Osteostatin; Parathyroid Hormone Like Hormone

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  • ELISA Kit for Parathyroid Hormone Related Protein (PTHrP) Packages (Simulation)
  • ELISA Kit for Parathyroid Hormone Related Protein (PTHrP) Packages (Simulation)
  • ELISA Kit for Parathyroid Hormone Related Protein (PTHrP) Results demonstration
  • CEA819Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Parathyroid Hormone Related Protein (PTHrP) and the recovery rates were calculated by comparing the measured value to the expected amount of Parathyroid Hormone Related Protein (PTHrP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-102 96
EDTA plasma(n=5) 96-103 101
heparin plasma(n=5) 80-92 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Parathyroid Hormone Related Protein (PTHrP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Parathyroid Hormone Related Protein (PTHrP) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Parathyroid Hormone Related Protein (PTHrP) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 80-103% 79-93% 78-98% 80-99%
EDTA plasma(n=5) 89-96% 83-97% 78-94% 78-94%
heparin plasma(n=5) 78-103% 96-105% 87-95% 79-96%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

ELISA Kit for Parathyroid Hormone Related Protein (PTHrP)

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. An antibody specific to PTHrP has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled PTHrP and unlabeled PTHrP (Standards or samples) with the pre-coated antibody specific to PTHrP. After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of PTHrP in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of PTHrP in the sample.

Citations

  • Characterization of a new renal cell carcinoma bone metastasis mouse modelPubMed: 20443133
  • Temporal activation of β-catenin signaling in the chondrogenic process of mesenchymal stem cells affects the phenotype of the cartilage generated.PubMed: 22133004
  • PTHrP Expression in Human MDA-MB-231 Breast Cancer Cells Is Critical for Tumor Growth and Survival and Osteoblast InhibitionPubMed: PMC3753447
  • Tetrahydrofurofuran-type lignans inhibit breast cancer-mediated bone destruction by blocking the vicious cycle between cancer cells, osteoblasts and osteoclastsPubmed: 23673814
  • Betulinic acid, a bioactive pentacyclic triterpenoid, inhibits skeletal-related events induced by breast cancer bone metastases and treatmentPubmed:24463094
  • Calcitonin gene-and parathyroid hormone-related peptides in normotensive and preeclamptic pregnancies: a nested case–control studyPubmed:24943060
  • 15-Deoxy-Δ12, 14-Prostaglandin J2 Inhibits Osteolytic Breast Cancer Bone Metastasis and Estrogen Deficiency-Induced Bone LossPubMed: 25859665
  • A role for the calcitonin receptor to limit bone loss during lactation in female mice by inhibiting osteocytic osteolysisPubMed: 26135836
  • Osteopontin expression in co-cultures of human squamous cell carcinoma-derived cells and osteoblastic cells and its effects on the neoplastic cell phenotype and osteoclastic activation.Pubmed:27312487
  • Hedgehog signaling in bone regulates whole-body energy metabolism through a bone–adipose endocrine relay mediated by PTHrP and adiponectinpubmed:27740628
  • Lactation induces increases in the RANK/RANKL/OPG system in maxillary bonePubmed:29382610
  • A Diet Rich in Fish Oil and Leucine Ameliorates Hypercalcemia in Tumour-Induced Cachectic MicePubmed: 31600911
  • Identification of a potential gene target for osteoarthritis based on bioinformatics analysesPubmed: 32571421
  • Parathyroid hormone related protein (PTHrP)-mediated hypercalcemia in malignancy associated with anti-PD-1 immune checkpoint inhibitor treatment and related …
  • Паратиреоидный гормон-родственный протеин как предиктор развития мочекаменной болезни
  • Extracellular vesicles-released parathyroid hormone-related protein from Lewis lung carcinoma induces lipolysis and adipose tissue browning in cancer ¡­33510128
  • Effects of whole-body vibration training in a cachectic C26 mouse model34732809
  • The acute adaptation of renal phosphate transporters in the murine kidney to oral phosphate intake requires multiple signalsPubmed:35334154

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