Mouse Cardiovascular ELISA Kits
Mouse ENG (Endoglin) CLIA Kit (MOES00532)
- SKU:
- MOES00532
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 9.38pg/mL
- Range:
- 15.63-1000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- CD105, END, HHT1, ORW, ORW1
- Reactivity:
- Mouse
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cardiovascular
Description
Assay type: | Sandwich |
Format: | 96T |
Assay time: | 4.5h |
Reactivity: | Mouse |
Detection method: | Chemiluminescence |
Detection range: | 15.63-1000 pg/mL |
Sensitivity: | 9.38 pg/mL |
Sample volume: | 100µL |
Sample type: | Serum, plasma and other biological fluids |
Repeatability: | CV < 15% |
Specificity: | This kit recognizes Mouse ENG in samples. No significant cross-reactivity or interference between Mouse ENG and analogues was observed. |
This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Mouse ENG. Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse ENG 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 Mouse ENG, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Mouse ENG. The concentration of Mouse ENG in the samples can be calculated by comparing the RLU of the samples to the standard curve.
UniProt Protein Function: | ENG: Major glycoprotein of vascular endothelium. May play a critical role in the binding of endothelial cells to integrins and/or other RGD receptors. Homodimer that forms an heteromeric complex with the signaling receptors for transforming growth factor-beta: TGFBR1 and/or TGFBR2. It is able to bind TGF-beta 1, and 3 efficiently and TGF-beta 2 less efficiently. Interacts with TCTEX1D4. Interacts with ARRB2. Endoglin is restricted to endothelial cells in all tissues except bone marrow. 2 isoforms of the human protein are produced by alternative splicing. |
UniProt Protein Details: | Protein type:Membrane protein, integral; Motility/polarity/chemotaxis; Receptor, misc. Chromosomal Location of Human Ortholog: 2 B|2 22. 09 cM Cellular Component: cell surface; cytoplasm; external side of plasma membrane; extracellular space; focal adhesion; nucleoplasm; receptor complex Molecular Function:galactose binding; glycosaminoglycan binding; protein binding; protein homodimerization activity; punt binding; transforming growth factor beta binding; transforming growth factor beta receptor, cytoplasmic mediator activity Biological Process: angiogenesis; artery morphogenesis; atrial cardiac muscle morphogenesis; central nervous system vasculogenesis; chronological cell aging; heart development; heart looping; negative regulation of cell migration; negative regulation of protein amino acid autophosphorylation; patterning of blood vessels; positive regulation of angiogenesis; positive regulation of BMP signaling pathway; positive regulation of collagen biosynthetic process; positive regulation of protein amino acid phosphorylation; positive regulation of transcription from RNA polymerase II promoter; regulation of transcription, DNA-dependent; regulation of transforming growth factor beta receptor signaling pathway; smooth muscle development; sprouting angiogenesis; transforming growth factor beta receptor signaling pathway; vasculogenesis; venous blood vessel morphogenesis |
UniProt Code: | Q63961 |
NCBI GenInfo Identifier: | 341940479 |
NCBI Gene ID: | 13805 |
NCBI Accession: | Q63961. 2 |
UniProt Secondary Accession: | Q63961,Q61520, Q8K100, |
UniProt Related Accession: | Q63961 |
Molecular Weight: | 70,021 Da |
NCBI Full Name: | Endoglin |
NCBI Synonym Full Names: | endoglin |
NCBI Official Symbol: | Eng |
NCBI Official Synonym Symbols: | Endo; CD105; AI528660; AI662476; S-endoglin |
NCBI Protein Information: | endoglin |
UniProt Protein Name: | Endoglin |
UniProt Synonym Protein Names: | Cell surface MJ7/18 antigen; CD_antigen: CD105 |
UniProt Gene Name: | Eng |
As the RLU 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.
Concentration (pg/mL) | RLU | Average | Corrected |
1000 | 50943 56407 | 53675 | 53650 |
500 | 22222 22756 | 22489 | 22464 |
250 | 11113 9309 | 10211 | 10186 |
125 | 4567 5235 | 4901 | 4876 |
62.5 | 2624 2282 | 2453 | 2428 |
31.25 | 1289 1273 | 1281 | 1256 |
15.63 | 674 742 | 708 | 683 |
0 | 24 26 | 25 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse ENG were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse ENG were tested on 3 different plates, 20 replicates in each plate.
Intra-assay Precision | Inter-assay Precision | |||||
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 20 | 20 | 20 |
Mean (pg/mL) | 46.16 | 110.62 | 373.66 | 46.71 | 117.42 | 340.95 |
Standard deviation | 5.78 | 9.41 | 25.00 | 5.04 | 8.99 | 34.78 |
C V (%) | 12.52 | 8.51 | 6.69 | 10.79 | 7.66 | 10.20 |
Recovery
The recovery of Mouse ENG spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
Sample Type | Range (%) | Average Recovery (%) |
Serum (n=5) | 91-104 | 98 |
EDTA plasma (n=5) | 97-111 | 103 |
Cell culture media (n=5) | 86-100 | 92 |
Linearity
Samples were spiked with high concentrations of Mouse ENG 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) | ||
1:2 | Range (%) | 92-104 | 95-113 | 88-100 |
Average (%) | 98 | 103 | 93 | |
1:4 | Range (%) | 101-117 | 87-99 | 86-96 |
Average (%) | 108 | 92 | 91 | |
1:8 | Range (%) | 98-115 | 100-118 | 90-105 |
Average (%) | 105 | 109 | 96 | |
1:16 | Range (%) | 90-105 | 93-107 | 95-107 |
Average (%) | 96 | 99 | 101 |
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.
Item | Specifications | Storage |
Micro CLIA 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 A | 1 vial, 5 mL | 4°C (shading light) |
Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
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 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
- Determine the RLU value of each well immediately.