Human Bone morphogenetic protein 2,BMP-2 ELISA Kit

Instructions
Code CSB-E04507h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The product CSB-E04507h manufactured by CUSABIO is a solid-phase sandwich ELISA kit that allows for the quantitative determination of bone morphogenetic protein 2 (BMP2) in the samples, including serum, urine, cell culture supernates, and tissue homogenates. This human BMP2 ELISA kit specifically recognizes the human BMP2. It has been quality validated through multiple criteria, including sensitivity, specificity, precision, recovery, and lot-to-lot consistency. More details are described in the product instructions.

Samples and standards are respectively pipetted into the appropriate micro ELISA plate wells pre-coated with anti-human BMP2 antibody. The Biotin-labeled BMP2 antibody is added to form a sandwich of immune complexes. HRP-avidin and TMB substrate are successively added, and the solution turns blue. The blue turns yellow after the addition of the stop solution. The color intensity is proportional to the AMH content in the sample.

BMP2 belongs to the TGF-β superfamily of proteins. It is involved in various biological processes, including bone and cartilage formation, tissue and organ development, and the modulation of cell differentiation, proliferation, angiogenesis, morphogenesis, chemotaxis, cellular survival, and apoptosis. BMP2 in cancer either exerts an inhibitive effect or acts as a tumor promoter. It can activate oncogenes and initiate metastasis progression in the tumor microenvironment. Gui Wu etc showed that high BMP2 concentrations augmented the expansion of myeloid-derived suppressor cells (MDSCs), which secreted IL-6, further enhancing liver cancer cell proliferation. BMP2 could suppress the proliferation of various cancer cell lines such as MDA-MB-231 breast cancer cells, osteosarcoma cells, and gastric cancer cells.

Target Name bone morphogenetic protein 2
Alternative Names BDA2 ELISA Kit; BMP-2 ELISA Kit; BMP-2A ELISA Kit; Bmp2 ELISA Kit; BMP2_HUMAN ELISA Kit; BMP2A ELISA Kit; Bone morphogenetic protein 2 ELISA Kit; Bone morphogenetic protein 2A ELISA Kit
Abbreviation BMP2
Uniprot No. P12643
Species Homo sapiens (Human)
Sample Types serum, urine, cell culture supernates, tissue homogenates
Detection Range 62.5 pg/mL-4000 pg/mL
Sensitivity 15.6 pg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Signal Transduction
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 BMP-2 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:1 Average % 90  
Range % 80-95  
1:2 Average % 95  
Range % 91-99  
1:4 Average % 97  
Range % 92-106  
1:8 Average % 93  
Range % 86-99  
Recovery
The recovery of human BMP-2 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 Type Average % Recovery Range  
Serum (n=5) 95 89-100  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
pg/ml OD1 OD2 Average Corrected  
4000 2.673 2.786 2.730 2.572  
2000 2.345 2.324 2.335 2.177  
1000 1.639 1.740 1.690 1.532  
500 1.180 1.200 1.190 1.032  
250 0.831 0.879 0.855 0.697  
125 0.592 0.605 0.599 0.441  
62.5 0.345 0.356 0.351 0.193  
0 0.154 0.162 0.158    
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human BMP2 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 BMP2 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.
  • 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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 Q&A
Q:

If the conformation of BMP-2 changes, is there a possibility that quantification of BMP-2 can not be done?

A:
Thanks for your inquiry.
About the change of conformation of BMP-2, may I know what causes that change?
Theoretically, we should not change conformation of the protein. Usually the change you mentioned is the change of the advanced conformation. For elisa assay, the detection target can be recognized as long as the recognition site of antibodies is not affected.
For CSB-E04507h,Human Bone morphogenetic protein 2,BMP-2 ELISA Kit, BMP-2 can be detected out if the concentration falls in the detection range of the kit.

Target Background

Function
(From Uniprot)
Induces cartilage and bone formation
Gene References into Functions
  1. These results not only call into question the use of VEGFA alone in bone regeneration, but also highlight the importance in BTE of appropriately formulated combined delivery of VEGFA and BMP2. PMID: 29386057
  2. This work indicates that NELL-1, HMGB1, and CCN2 might enhance bone defect healing via the recruitment of endogenous cells and induction of vascularization and act via different processes than BMP2. PMID: 28463604
  3. Serum BMP2 and Smad4 levels in patients with senile osteoporotic fracture were significantly lower than those in normal controls PMID: 29938690
  4. conclude that SUMO3-tagged hBMP2 is more suited for generation of soluble form of the protein and addition of SUMO3 tag does not affect the functional activity of hBMP2 PMID: 29574511
  5. The present study identified a change in miR-22, miR-140, and BMP-2 expression in the synovial fluid of patients with osteoarthritis before and after arthroscopic debridement. PMID: 29429984
  6. The study revealed an enhanced sensitivity of aortic valve interstitial cells to osteogenic inductors in aortic stenosis patients, which indicates probable implication of OPN, OPG, and BMP2 genes in pathogenesis of aortic valve calcification. PMID: 29308559
  7. The rhBMP2 monomer and dimer were eluted at 0.9 M and 0.6 M NaCl, respectively. The alkaline phosphatase assay of rhBMP2 (0, 50, 100, 200, and 400 ng/ml) was analyzed on C2C12 cells and maximum 200 ng/ml activity was observed in dose dependent manner PMID: 29333457
  8. In contrast to BMP-2, BMP-7 concomitantly inhibited the expression of profibrotic genes PMID: 28102712
  9. The binding free energies indicate that ALK-3 preferably binds to BMP-2 instead of BMP-9. The structural analysis shows that ALK-3 binding with BMP-2 occurs in a perfectly symmetry pathway, whereas this symmetry is lost for possible ALK-3 interactions with BMP-9 PMID: 28869862
  10. The results demonstrate the efficacy of HPP-GC hydrogel in minimizing the diffusive loss of rhBMP-2 from the implantation site, compared to the collagen hydroxyapatite scaffold. PMID: 28847606
  11. The in vitro results suggest that altered BMP2 regulatory function at rs1884302 may contribute to the etiology of sagittal nonsyndromic craniosynostosis. The in vivo results indicate that differences in regulatory activity depend on the presence of a C or T allele at rs1884302. PMID: 28985029
  12. Collectively, according to our study, rhIL-6 could induce the extracellular calcification and osteogenic differentiation of human artery smooth muscle cells through upregulating endogenous BMP2 in vitro. This may be one of the underlying mechanisms of the overwhelming vascular calcification in rheumartoid arthritis. PMID: 28134597
  13. HUCB-MSC transfected with mTAT/PEI were shown to express more BMP-2 protein and mRNA. PMID: 28951869
  14. These results showed that BMP2 activated SMAD1/5/8 phosphorylation and up-regulated BAMBI mRNA in human granulosa-lutein cells. PMID: 28578012
  15. BMP-2 can enhance HUVEC proliferation, migration and angiogenesis through P38, ERK and Akt/m-TOR pathway. PMID: 27886213
  16. Study shows that recombinant human bone morphogenetic protein-2 activates hippo signaling through RASSF1 in esophageal cancer cells PMID: 27230238
  17. SNPs in BMP2 can predict grade >/= 2 or 3 RP after radiotherapy for NSCLC and improve the predictive power of MLD model. PMID: 28574846
  18. CTGF and BMP2 are induced following myocardial ischemia in mice and humans. PMID: 28460577
  19. Missense mutations in COL6A1, COL11A2, FGFR1, and BMP2 genetically predispose patients to ossification of posterior longitudinal ligaments. PMID: 27246988
  20. Computational analysis on conformational dynamics of BMP-2 has been presented. PMID: 27426435
  21. there was a significant association in men between BMP2 genetic variant (rs235756) and hypertension in the genetically homogeneous Finnish population; no significant association between BMP2 rs235768 (A>T) and hypertension was found PMID: 29390526
  22. Adding NMP as an adjunct to rhBMP-2-coated BCP produced inconsistent effects on bone regeneration, resulting in no significant benefit compared to controls. PMID: 28680881
  23. observations regarding the dysregulation of these gatekeepers of neuronal viability may have important implications in understanding the iAbeta1-42 mediated effects observed in AD PMID: 29470488
  24. This study demonstrates that viscous collagen gel can be an effective carrier for rhBMP-2 delivery into surgical sites, and that the injectable rhBMP-2-containing collagen gel may be applied for the enhancement of tendon-bone interface healing PMID: 26177709
  25. Synergistic effects of BMP-2, BMP-6 or BMP-7 with human plasma fibronectin onto hydroxyapatite coatings. PMID: 28434979
  26. High-dose recombinant human bone morphogenetic protein-2 impacts histological and biomechanical properties of a cervical spine fusion segment: results from a sheep model. PMID: 26053675
  27. Report osteoblast-like transformation of epithelial breast cancer cells that have undergone epithelial-mesenchymal transition followed by bone morphogenetic protein-2 stimulation. RUNX2 functions as a master mediator of this process. PMID: 27806311
  28. Taken together, our results suggest that DHCA may be developed as an efficient therapeutic for osteoporosis by regulating osteoblastogenesis through its estrogenic effects. PMID: 29253565
  29. BMP2-transduced BMSCs can maintain the chondrocyte-like phenotype in PRP gel in vitro, and the combined use of these two agents can significantly promote repair of the degenerated discs in vivo PMID: 26169838
  30. these results suggest that the BMP2 gene polymorphism may be related to the development of allograft rejection and graft dysfunction in kidney transplant recipients PMID: 28583517
  31. Data show that the GREMLIN 2 (GREM2) expression during Induced Pluripotent Stem Cell (hiPS) cell cardiac differentiation follows the expression pattern of cardiac-specific genes. PMID: 28125926
  32. Results identify a novel 4671-bp tandem duplication downstream of BMP2, which is associated with brachydactyly type A2 . The duplication highly overlaps the sequences reported previously but has a different breakpoint and a different flanking microhomology. PMID: 29129813
  33. miR-106b inhibited osteoblastic differentiation and bone formation partly through directly targeting bone morphogenetic protein 2. PMID: 28108317
  34. BMP2 decreases gap junction intercellular communication of luteinized human granulosa cells by downregulating Cx43 expression through an ALK2/ALK3-mediated SMAD-dependent signaling pathway. PMID: 27986931
  35. BMP2 also requires Src for filamentous actin polymerization in Tgfbr3(-/-) epicardial cells. PMID: 26645362
  36. The deletion contained 17 protein coding genes including PROKR2 and BMP2, both of which are expressed during embryological development of the pituitary gland. PROKR2 mutations have been associated with hypopituitarism but a heterozygous deletion of this gene with hypopituitarism is a novel observation. PMID: 28586151
  37. both bone morphogenetic protein 2 (BMP2) and BMP6 are proangiogenic in vitro and ex vivo and that the BMP type I receptors, activin receptor-like kinase 3 (ALK3) and ALK2, play crucial and distinct roles in this process. PMID: 28733457
  38. sequential presentation of PDGF to BMP-2 led to increased tubule formation over simultaneous delivery of these growth factors. PMID: 27650131
  39. Bone Morphogenetic Protein-2, But Not Mesenchymal Stromal Cells, Exert Regenerative Effects on Canine and Human Nucleus Pulposus Cells PMID: 27829314
  40. The structure of Grem2-GDF5 complex has revealed a number of key findings for DAN-family mediated BMP2 inhibition. PMID: 27524626
  41. Bioluminescence imaging reveals increased MSC survival when implanted in BMP-2 PAHs. PMID: 27581621
  42. Bone morphogenetic protein 2 promotes osteogenesis of bone marrow stromal cells in type 2 diabetic rats via the Wnt signaling pathway PMID: 27702654
  43. monocytes interact specifically with Chitosan-Fibrinogen (Ch-Fg) via TLR-4, triggering particular intracellular signalling pathways (ERK and JNK, but not p38), downstream of TLR-4. Functionally, Ch-Fg induced monocytes to produce the osteogenic mediator BMP-2. PMID: 27856281
  44. This study showed that si-Grem2 increased the BMP-2-induced osteogenic differentiation of hBMSCs via the BMP-2/Smad/Runx2 pathway. PMID: 27335248
  45. Low doses of IL1B activate the BMP/Smad signaling pathway to promote the osteogenesis of periodontal ligament stem cells, but higher doses of IL1B inhibit BMP/Smad signaling through the activation of NF-kappaB and MAPK signaling, inhibiting osteogenesis. PMID: 27415426
  46. Increased miR-93-5p in trauma-induced osteonecrosis of the femoral head patients inhibited osteogenic differentiation, which may be associated with BMP-2 reduction. PMID: 28797104
  47. RANKL promotes VC by inducing BMP-2 release from HAECs PMID: 27339040
  48. KDM5A-mediated H3K4me3 modification participated in the etiology of osteoporosis and may provide new strategies to improve the clinical efficacy of BMP2 in osteoporotic conditions. PMID: 27512956
  49. fabricated scaffolds were well coated with DOPA as well as grafted with rhBMP2 at a quantity of 22.7+/-5ng when treatment with 100ng/ml rhBMP2 and 153.3+/-2.4ng when treated with 500ng/ml rhBMP2. This grafting enables rhBMP2 to be released in a sustained pattern. PMID: 26868173
  50. Data suggest pituitary cells secrete factor (TSP1) that binds to and inhibits action of BMP2 and BMP4; von Willebrand type C domain of TSP1 is likely responsible for this BMP2/4-binding activity. These studies were initially conducted using cultured cells from ovine pituitary gland and mouse cell line; interactions were confirmed using recombinant human proteins. (TSP1 = thrombospondin-1; BMP = bone morphogenetic protein) PMID: 28747434

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Subcellular Location Secreted
Protein Families TGF-beta family
Tissue Specificity Particularly abundant in lung, spleen and colon and in low but significant levels in heart, brain, placenta, liver, skeletal muscle, kidney, pancreas, prostate, ovary and small intestine.
Database Links

HGNC: 1069

OMIM: 112261

KEGG: hsa:650

STRING: 9606.ENSP00000368104

UniGene: Hs.73853

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