Human bone alkaline phosphatase,BALP ELISA Kit

Instructions
Code CSB-E09033h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human bone alkaline phosphatase (BALP) ELISA Kit quantitates BALP levels in human serum, plasma, or tissue homogenates. BALP is an enzyme that promotes bone mineralization by inactivating pyrophosphate and osteopontin, which both are inhibitors of bone mineralization. It is essential in determining the rate of bone formation and turnover. BALP is regarded as a marker of bone formation. Recent studies demonstrated BALP plays a crucial role in the pathogenesis of vascular calcification and identified BALP as a promising predictor of mortality in chronic kidney disease (CKD).

This kit employs the sandwich-ELISA mechanism in conjugation with BALP antibody-BALP antigen-specific binding as well as HRP-TMB chromogenic reaction to measure the concentration of BALP in the samples. The kit is catheterized with high sensitivity, strong specificity, good linearity, recovery of 89%-108%, precision less than 10%, and lot-to-lot consistency. Refer to the product instructions for more validation information.

Target Name bone alkaline phosphatase,BALP
Alternative Names AKP2 ELISA Kit; Alkaline phosphatase liver/bone/kidney ELISA Kit; Alkaline phosphatase liver/bone/kidney isozyme ELISA Kit; Alkaline phosphatase tissue nonspecific isozyme ELISA Kit; Alkaline phosphatase, tissue-nonspecific isozyme ELISA Kit; Alkaline phosphomonoesterase ELISA Kit; Alpl ELISA Kit; AP TNAP ELISA Kit; AP-TNAP ELISA Kit; APTNAP ELISA Kit; BAP ELISA Kit; FLJ40094 ELISA Kit; FLJ93059 ELISA Kit; Glycerophosphatase ELISA Kit; HOPS ELISA Kit; Liver/bone/kidney type alkaline phosphatase ELISA Kit; MGC161443 ELISA Kit; MGC167935 ELISA Kit; PHOA ELISA Kit; PPBT_HUMAN ELISA Kit; Tissue non specific alkaline phosphatase ELISA Kit; Tissue nonspecific ALP ELISA Kit; TNAP ELISA Kit; TNSALP ELISA Kit
Abbreviation BALP
Uniprot No. P05186
Species Homo sapiens (Human)
Sample Types serum, plasma, tissue homogenates
Detection Range 3.12 ng/mL-200 ng/mL
Sensitivity 0.78 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Cancer
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human BALP in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
 SampleSerum(n=4)
1:1Average %100
Range %96-107
1:2Average %89
Range %85-96
1:4Average %92
Range %86-98
1:8Average %93
Range %88-98
Recovery
The recovery of human BALP spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample TypeAverage % RecoveryRange
Serum (n=5) 9489-97
EDTA plasma (n=4)10297-108
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/mlOD1OD2AverageCorrected
2002.492 2.392 2.442 2.311
1001.912 1.812 1.862 1.731
501.283 1.183 1.233 1.102
250.699 0.679 0.689 0.558
12.50.426 0.416 0.421 0.290
6.250.334 0.324 0.329 0.198
3.120.218 0.208 0.213 0.082
00.132 0.129 0.131  
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human BALP antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard --- Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled BALP antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) --- Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody solution.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) --- Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) --- Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) --- Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

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Target Background

Function
(From Uniprot)
This isozyme may play a role in skeletal mineralization.
Gene References into Functions
  1. a significant proportion of adult heterozygotes for ALPL mutations may have unspecific symptoms not attributable to their heterozygosity. PMID: 29236161
  2. The ALPL SNP, rs1256328, was identified as being significantly associated with kidney stone disease status in a large Chinese Han cohort PMID: 29489416
  3. The expression of ALP mRNA and ALP activity in bone tissue were much higher in osteoporosis patients with fracture than those without fracture. PMID: 29786747
  4. genu varum is associated with the alkaline phosphatase level regardless of the presence of radiographic abnormalities in the growth plate in children. PMID: 28664247
  5. Adults with persistent hypophosphatasemia frequently harbor alkaline phosphatase mutations and have elevated ALP substrates. PMID: 28401263
  6. Mutations in ALPL which reduce alkaline phosphatase activity are responsible for Hypophosphatasia , a rare disorder characterized by defective bone and teeth mineralization and early tooth loss. PMID: 28570402
  7. These results show that an increase of TNAP activity in ACDC (arterial calcification due to deficiency of CD73) contributes to ectopic calcification by disrupting the extracellular balance of PPi and Pi and identify potential therapeutic targets for ACDC. PMID: 27965423
  8. the identification of 11 novel ALPL mutations in the five different HPP forms and the observation of a recurrent mutation, p. (Thr166Ile) in the Spanish population expand our knowledge of pathogenic ALPL mutations. PMID: 28127875
  9. Preoperative calcitonin levels were correlated with the presence of tumor, whereas alkaline phosphatase (ALP) levels were not. There were no significant associations between tumor volume and ALP or calcitonin levels in the preoperative or postoperative periods. During long-term follow-up, serum ALP was significantly associated with tumor recurrence, but serum calcitonin was not. PMID: 27922893
  10. His ALPL gene mutation came from c.228delG mutation in his mother and c.407G>A compound heterozygous mutation in his father PMID: 28506345
  11. Both PPARgamma gene expression and TNALP activity increased during intracellular lipid accumulation in HepG2 and 3T3-L1 cells. Inhibition of TNALP blocked intracellular lipid accumulation but did not alter expression of the PPARgamma gene. PMID: 28209522
  12. ALPL is a major contributor to the pathogenesis of Prostate cancer progression. PMID: 28006818
  13. This result indicated that the 1559delT mutant was not retained on the plasma membrane owing to a lack of the Glycosylphosphatidylinositol anchor. PMID: 27680481
  14. ALPL expression is significantly upregulated in human masticatory mucosa during wound healing PMID: 28005267
  15. serum ALP levels were not associated with increased death risk in prevalent HD patients over a 5-year interval. PMID: 27467278
  16. In conclusion, serum levels of BSP, ALP, ICTP, and PSA increased in patients with bone metastases, and combined detection of all markers could improve the positive-predictive value. PMID: 27323113
  17. results reveal that the amino acid substitutions at position 426 of TNSALP differentially affect the structure and function of TNSALP, leading to understanding of the molecular and cellular basis of hypophosphatasia. PMID: 28000043
  18. One-half of adult individuals with unexplained low serum ALP carried an ALPL mutation. The presence of a mutated allele was associated with tooth loss, slightly lower levels of serum ALP, higher levels of pyridoxal phosphate and phosphoethanolamine, as well as mildly increased serum phosphate. PMID: 26783040
  19. Dynamic changes of ALP, LDH and PSA during Abiraterone-therapy are associated with best clinical benefit and OS in bone metastatic castration resistant prostate cancer PMID: 26975660
  20. glycosylation differences in human bone alkaline phosphatases are of crucial importance for protein-protein interactions with collagen type I PMID: 26645431
  21. Analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. PMID: 26797772
  22. The presence of TNAP increased the dynamics and decreased the ordering of model membranes. PMID: 26389140
  23. These data confirm that TNAP is co-expressed by dental pulp stromal cells together with other bone marrow stromal cells markers and show that cell density affects TNAP expression levels. PMID: 25636587
  24. ALP quartiles were significantly associated with albuminuria in participants with estimated glomerular filtration rate>120 90-119, 60-89 and <60 mL/min/1.73 Higher ALP levels are significantly associated with renal hyperfiltration PMID: 25853240
  25. during skeletal mineralization, the building Ca2+ gradient first activates TNAP, but gradually inactivates it at high Ca2+ concentrations, toward completion of mineralization. PMID: 25775211
  26. these data demonstrate that TNAP activity is significantly increased in the brain in both the sporadic and familial forms of Alzheimer's Disease (AD) and that TNAP activity is significantly increased in the plasma in AD patients PMID: 26219720
  27. A non-linear relationship exists between serum levels of ALP and phosphate and risk of total mortality from cardiovascular diseases. PMID: 25033287
  28. Higher alkaline phosphatase was associated with the short-term adverse outcomes of peritoneal dialysis-related peritonitis PMID: 25246707
  29. ALP mRNA binds to and is stabilized by vimentin. PMID: 25536665
  30. Elevated AP was associated with the presence of COPD and respiratory symptoms (cough, wheezing). PMID: 25336462
  31. Two-month alkaline phosphatase of <100 U/L had a negative predictive value of 97% for development of ischemic cholangiopathy after liver transplantation. PMID: 25769592
  32. A novel role of alkaline phosphatase in the ERK1 and ERK2 dephosphorylation in renal cell carcinoma cell lines PMID: 25241253
  33. In inflammatory cholestatic conditions, loss of activity of liver AP might promote hyper-adenosine triphosphate-bilia, lipopolysaccharide overload, and subsequent exacerbation and perpetuation of inflammation. [review] PMID: 25603770
  34. patterns were confirmed in human teeth, including widespread TNAP, and NPP1 restricted to cementoblasts lining acellular cementum PMID: 25504209
  35. High levels of alkaline phosphatase (a biochemical markers of osteosynthesis) is associated with poor prognosis in metastatic bone cancer from disseminated breast cancer. PMID: 25342482
  36. Polymorphisms in ALP, ENPP1 and ANKH are important genetic risk factors contributing to Pseudoxanthoma elasticum PMID: 25025693
  37. REVIEW: role of bone-type tissue-nonspecific alkaline phosphatase and PHOSPO1 in vascular calcification PMID: 24533943
  38. This family report indicates that mapping ALPL mutations within the gene does not necessarily help to predict the clinical severity of the hypophosphatasia phenotype. PMID: 24569605
  39. Data indicate that alkaline phosphatase (AP) velocity kinetics (APV)is an independent predictor of overall survival (OS) and bone metastasis-free survival (BMFS) in patients with -resistant prostate cancer (CRPC). PMID: 24929891
  40. the new role of ALP in cell viability and apoptosis and involvement in renal cell carcinoma tumorigenesis PMID: 24909115
  41. Effect of cyclic mechanical stimulation on the expression of osteogenesis genes in human intraoral mesenchymal stromal and progenitor cells. PMID: 24804200
  42. DNMT inhibitors facilitate the Pi-induced development of vascular calcification via the upregulation of the ALP expression along with a reduction in the DNA methylation level of the ALP promoter region. PMID: 24441913
  43. ABO locus is a major determinant for serum ALP levels in Chinese Han population. PMID: 24094242
  44. The CPT score,alkaline phosphatase > 1.5 ULN, and the CS nonresponse had an independent impact on the 90-day survival in alcoholic hepatitis. PMID: 24151614
  45. characterization of a novel genetic alteration (c.1318_1320delAAC, p.N440del) in the ALPL gene resulting in odonto-hypophosphatasia(HPP) in monozygotic twins; results assist in defining genotype-phenotype associations for odonto-HPP and identify the collagen-binding site as a region of potential structural importance for TNAP function in the biomineralization PMID: 23791648
  46. Serum ALP is adversely associated with measures of arterial structure and function in hypertensive African men. PMID: 22656046
  47. The Y28D, A111T and T389N mutants displayed only negligible ALP activity in vitro compared to the wild-type (WT) tissue-nonspecific alkaline phosphatase. PMID: 24022022
  48. The most frequent clinical type was the PLH type with prognosis related to respiratory failure, biochemical/radiological changes and ALPL mutations. PMID: 24276437
  49. The aim of this study was to investigate two mineralization-related genes TNAP and ANKH polymorphisms associated with ankylosing spondylitis (AS) in the North Chinese Han population. PMID: 23612078
  50. data suggest that the promineralization role of TNAP may be related not only to its accepted pyrophosphatase activity but also to its ability to modify the phosphorylation status of OPN. PMID: 23427088

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Involvement in disease Hypophosphatasia (HOPS); Hypophosphatasia childhood type (HOPSC); Hypophosphatasia infantile type (HOPSI)
Subcellular Location Cell membrane, Lipid-anchor, GPI-anchor
Protein Families Alkaline phosphatase family
Database Links

HGNC: 438

OMIM: 146300

KEGG: hsa:249

STRING: 9606.ENSP00000363965

UniGene: Hs.75431

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