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Human GDF6 (Growth Differentiation Factor 6) ELISA Kit (HUES02858)

SKU:
HUES02858
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q6KF10
Sensitivity:
0.09ng/mL
Range:
0.16-10ng/mL
ELISA Type:
Sandwich
Synonyms:
BMP13, CDMP2, KFS, KFSL, MCOP4, MGC158100, MGC158101, SCDO4, SGM1,MCOPCB6,KFS1,
Reactivity:
Human
Sample Type:
Serum, plasma and other biological fluids
Research Area:
Cell Death
€599
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Description

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Human GDF6 (Growth Differentiation Factor 6) ELISA Kit

The Human GDF6 (Growth Differentiation Factor 6) ELISA Kit is a powerful tool for the accurate measurement of GDF6 levels in human samples, including serum, plasma, and cell culture supernatants. With high sensitivity and specificity, this kit delivers reliable and reproducible results for a variety of research applications.GDF6 is a key factor in bone and cartilage development, playing a crucial role in skeletal growth and repair. Its dysregulation has been linked to various skeletal disorders, making it a valuable biomarker for studying these conditions and potentially developing targeted therapies.

This ELISA kit offers researchers a precise and efficient way to quantify GDF6 levels, providing valuable insights into its role in skeletal biology and related pathologies. Trust in the Human GDF6 ELISA Kit for accurate and dependable results in your research endeavors.

Assay type:Sandwich
Format:96T
Assay time:4.5h
Reactivity:Human
Detection Method:Colormetric
Detection Range:0.16-10 ng/mL
Sensitivity:0.10 ng/mL
Sample Volume Required Per Well:100µL
Sample Type:Serum, plasma and other biological fluids
Specificity:This kit recognizes Human GDF6 in samples. No significant cross-reactivity or interference between Human GDF6 and analogues was observed.

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human GDF6. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human GDF6 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 Human GDF6, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human GDF6. The concentration of Human GDF6 in samples can be calculated by comparing the OD of the samples to the standard curve.

UniProt Protein Function:GDF6: Required for normal formation of bones and joints in the limbs, skull, and axial skeleton. Plays a key role in establishing boundaries between skeletal elements during development. Defects in GDF6 are the cause of Klippel-Feil syndrome type 1 (KFS1). A skeletal disorder characterized by congenital fusion of cervical vertebrae. It is due to a failure in the normal segmentation of vertebrae during the early weeks of fetal development. The clinical triad consists of short neck, low posterior hairline, and limited neck movement. Deafness is a well- known feature of KFS and may be of sensorineural, conductive, or mixed type. A chromosomal aberration involving GDF6 has been found in a patient with Klippel-Feil syndrome (KFS). Paracentric inv(8)(q22;2q23. 3). Defects in GDF6 are the cause of microphthalmia isolated type 4 (MCOP4). A disorder of eye formation, ranging from small size of a single eye to complete bilateral absence of ocular tissues. Ocular abnormalities like opacities of the cornea and lens, scaring of the retina and choroid, cataract and other abnormalities like cataract may also be present. Belongs to the TGF-beta family.
UniProt Protein Details:

Protein type:Cytokine; Cell development/differentiation; Secreted, signal peptide; Secreted

Chromosomal Location of Human Ortholog: 8q22. 1

Cellular Component: extracellular space

Molecular Function:cytokine activity; growth factor activity; protein homodimerization activity; transforming growth factor beta receptor binding

Biological Process: activin receptor signaling pathway; apoptosis; BMP signaling pathway; cell development; growth; positive regulation of neuron differentiation; positive regulation of transcription, DNA-dependent; regulation of apoptosis; regulation of MAPKKK cascade

Disease: Klippel-feil Syndrome 1, Autosomal Dominant; Leber Congenital Amaurosis 17; Microphthalmia, Isolated 4; Microphthalmia, Isolated, With Coloboma 6

NCBI Summary:This gene encodes a member of the bone morphogenetic protein (BMP) family and the TGF-beta superfamily of secreted signaling molecules. It is required for normal formation of some bones and joints in the limbs, skull, and axial skeleton. Mutations in this gene result in colobomata, which are congenital abnormalities in ocular development, and in Klippel-Feil syndrome (KFS), which is a congenital disorder of spinal segmentation. [provided by RefSeq, Jul 2008]
UniProt Code:Q6KF10
NCBI GenInfo Identifier:74748876
NCBI Gene ID:392255
NCBI Accession:Q6KF10. 1
UniProt Secondary Accession:Q6KF10,Q6PI58,
UniProt Related Accession:Q6KF10
Molecular Weight:50,662 Da
NCBI Full Name:Growth/differentiation factor 6
NCBI Synonym Full Names:growth differentiation factor 6
NCBI Official Symbol:GDF6
NCBI Official Synonym Symbols:KFM; KFS; KFS1; KFSL; SGM1; BMP13; CDMP2; LCA17; MCOP4; SCDO4; BMP-13; MCOPCB6
NCBI Protein Information:growth/differentiation factor 6
UniProt Protein Name:Growth/differentiation factor 6
UniProt Synonym Protein Names:Bone morphogenetic protein 13; BMP-13; Growth/differentiation factor 16
Protein Family:Growth/differentiation factor
UniProt Gene Name:GDF6
UniProt Entry Name:GDF6_HUMAN

As the OD 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
(ng/mL)		O.DAverageCorrected
102.221
2.261		2.2412.17
51.439
1.479		1.4591.388
2.50.824
0.8		0.8120.741
1.250.433
0.451		0.4420.371
0.630.211
0.211		0.2110.14
0.320.17
0.154		0.1620.091
0.160.117
0.119		0.1180.047
00.071
0.071		0.071--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level HumanGDF6 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level HumanGDF6 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay PrecisionInter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.531.163.780.571.123.69
Standard deviation0.030.050.120.030.060.13
C V (%)5.664.313.175.265.363.52

Recovery

The recovery of HumanGDF6 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample TypeRange (%)Average Recovery (%)
Serum (n=5)89-10094
EDTA plasma (n=5)92-10599
Cell culture media (n=5)85-10091

Linearity

Samples were spiked with high concentrations of HumanGDF6 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)
1:2Range (%)86-10193-10896-108
Average (%)9299101
1:4Range (%)89-10380-9085-97
Average (%)958590
1:8Range (%)87-10281-9683-94
Average (%)948788
1:16Range (%)86-9982-9288-101
Average (%)928795

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 ELISA 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 1 vial, 10 mL 4°C(shading light)
Stop Solution 1 vial, 10 mL 4°C
Plate Sealer 5 pieces
Product Description 1 copy
Certificate of Analysis 1 copy
  1. 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.
  2. Aliquot 100µl of standard solutions into the standard wells.
  3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
  4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
  5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
  6. 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.
  7. 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.
  8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
  9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
  10. Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
  11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
  12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

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