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Mouse Bone morphogenetic protein 5 (Bmp5) ELISA Kit (MOEB2410)

SKU:
MOEB2410
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P49003
ELISA Type:
Sandwich
Reactivity:
Mouse
€649
Frequently bought together:

Description

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Mouse Bone morphogenetic protein 5 (Bmp5) ELISA Kit

The Mouse Bone Morphogenetic Protein 5 (BMP5) ELISA Kit is a powerful tool for measuring levels of BMP5 in mouse serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit delivers accurate and reliable results, making it suitable for a variety of research applications.BMP5 is a key growth factor that plays a critical role in skeletal development and regeneration. It is involved in bone formation and influences the differentiation of mesenchymal stem cells into bone-forming osteoblasts.

Understanding the levels of BMP5 can provide valuable insights into skeletal disorders, fractures, and bone healing processes.This ELISA kit offers researchers a precise method for quantifying BMP5 levels, allowing for the identification of potential biomarkers and the development of targeted therapies for bone-related conditions. Trust in the Mouse BMP5 ELISA Kit for robust and trustworthy results in your research endeavors.

Product Name:Mouse Bone morphogenetic protein 5 (Bmp5) ELISA Kit
SKU:MOEB2410
Size:96T
Target:Mouse Bone morphogenetic protein 5 (Bmp5)
Synonyms:BMP-5, Bmp-5
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Mouse
Detection Range:0.312-20ng/mL
Sensitivity:0.183ng/mL
Intra CV:4.6%
Inter CV:6.4%
Linearity:
Sample1:21:41:81:16
Serum(N=5)94-107%82-92%82-92%89-100%
EDTA Plasma(N=5)103-113%97-107%81-90%103-113%
Heparin Plasma(N=5)102-111%108-117%100-110%90-100%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum9589-101
Plasma9791-103
Function:Induces cartilage and bone formation.
Uniprot:P49003
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant mouse Bone morphogenetic protein 5
Sub Unit:Homodimer; disulfide-linked.
Research Area:Cancer
Subcellular Location:Secreted
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:BMP5: Induces cartilage and bone formation. Belongs to the TGF-beta family.Protein type: Motility/polarity/chemotaxis; Secreted; Secreted, signal peptideChromosomal Location of Human Ortholog: 9 D|9 42.34 cMCellular Component: extracellular region; extracellular space; vesicleMolecular Function: cytokine activity; growth factor activity; protein binding; transforming growth factor beta receptor bindingBiological Process: BMP signaling pathway; cardiac muscle tissue development; cartilage development; cell development; cell differentiation; hindbrain development; male genitalia development; multicellular organism development; negative regulation of aldosterone biosynthetic process; negative regulation of cell proliferation; negative regulation of insulin-like growth factor receptor signaling pathway; neural fold elevation formation; ossification; pattern specification process; pharyngeal system development; positive regulation of epithelial cell proliferation; positive regulation of transcription from RNA polymerase II promoter; regulation of apoptosis; regulation of MAPK cascade; skeletal system development
UniProt Protein Details:
NCBI Summary:This gene encodes a secreted ligand of the TGF-beta (transforming growth factor-beta) superfamily of proteins. Ligands of this family bind various TGF-beta receptors leading to recruitment and activation of SMAD family transcription factors that regulate gene expression. The encoded preproprotein is proteolytically processed to generate each subunit of the disulfide-linked homodimer, which plays a role in bone and cartilage development. Mice with null mutations in this gene exhibit a short ear phenotype, which is characterized by reduced size of the external ear, altered size and shape of the sternum, and other skeletal and soft-tissue abnormalities. [provided by RefSeq, Jul 2016]
UniProt Code:P49003
NCBI GenInfo Identifier:66792782
NCBI Gene ID:12160
NCBI Accession:NP_031581.2
UniProt Secondary Accession:P49003,Q8CCE0
UniProt Related Accession:P49003
Molecular Weight:51,728 Da
NCBI Full Name:bone morphogenetic protein 5 preproprotein
NCBI Synonym Full Names:bone morphogenetic protein 5
NCBI Official Symbol:Bmp5
NCBI Official Synonym Symbols:se; AU023399
NCBI Protein Information:bone morphogenetic protein 5
UniProt Protein Name:Bone morphogenetic protein 5
UniProt Synonym Protein Names:
Protein Family:Bone morphogenetic protein
UniProt Gene Name:Bmp5
UniProt Entry Name:
Component Quantity (96 Assays) Storage
ELISA Microplate (Dismountable) 8×12 strips -20°C
Lyophilized Standard 2 -20°C
Sample Diluent 20ml -20°C
Assay Diluent A 10mL -20°C
Assay Diluent B 10mL -20°C
Detection Reagent A 120µL -20°C
Detection Reagent B 120µL -20°C
Wash Buffer 30mL 4°C
Substrate 10mL 4°C
Stop Solution 10mL 4°C
Plate Sealer 5 -

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir

*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.

Step
1. Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2. Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3. Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5. Repeat the wash process for five times as conducted in step 3.
6. Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7. Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8. Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9. After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol
Serum If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.
Plasma Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal Fluid Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatant Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysates Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenates The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysates Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast Milk Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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