Human TERF1 (Telomeric Repeat Binding Factor 1) ELISA Kit
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 TERF1 . 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 TERF1 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 TERF1, 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 TERF1. The concentration of Human TERF1 in samples can be calculated by comparing the OD of the samples to the standard curve.
|Detection Range||0.16-10 ng/mL|
|Sample Volume Required Per Well||100uL|
|Sample Type||Serum, plasma and other biological fluids|
This kit recognizes Human TERF1 in samples. No significant cross-reactivity or interference between Human TERF1 and analogues was observed.
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.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human TERF1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human TERF1 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||6.12||4.63||4.35||6.12||5.15||5.36|
The recovery of Human TERF1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||91-105||98|
|Cell culture media (n=5)||90-101||96|
Samples were spiked with high concentrations of Human TERF1 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)|
Kit Components & Storage
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.
|Micro ELISA Plate(Dismountable)||8 wells X12 strips||-20'C, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100X)||1 vial, 120 uL|
|Concentrated HRP Conjugate (100X)||1 vial, 120 uL||-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 (25X)||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|
Human TERF1 (Telomeric Repeat Binding Factor 1) ELISA Kit (HUES02951) Assay procedure
- 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.
Human TERF1 (Telomeric Repeat Binding Factor 1) ELISA Kit (HUES02951) Protein Information
|UniProt Protein Function:||TRF1: a telomeric repeat binding factor. Binds the telomeric double-stranded TTAGGG repeat and negatively regulates telomere length. Colocalizes with telomeric DNA in interphase and metaphase cells and is located at chromosome ends during metaphase. Involved in the regulation of the mitotic spindle. Expression is tightly regulated during the cell cycle; levels are low in G1 and S phase and increase during G2 phase and mitosis. Two splice variant isoforms have been described.|
|UniProt Protein Details:|
Chromosomal Location of Human Ortholog: 8q21.11
Cellular Component: nucleoplasm; chromosome, telomeric region; nuclear telomere cap complex; nuclear chromosome, telomeric region; cytoplasm; nucleolus; spindle; nucleus
Molecular Function:protein binding; protein homodimerization activity; DNA binding; protein heterodimerization activity; microtubule binding; telomeric DNA binding; double-stranded telomeric DNA binding; chromatin binding; DNA bending activity
Biological Process: response to drug; mitosis; positive regulation of mitosis; negative regulation of telomere maintenance via semi-conservative replication; positive regulation of apoptosis; positive regulation of microtubule polymerization; telomere maintenance via telomerase; positive regulation of mitotic cell cycle; negative regulation of telomerase activity; cell division; negative regulation of DNA replication; telomeric loop formation; mitotic cell cycle spindle assembly checkpoint; age-dependent telomere shortening; G2/M transition of mitotic cell cycle; negative regulation of telomere maintenance via telomerase; telomere maintenance; protein homooligomerization
|NCBI Summary:||This gene encodes a telomere specific protein which is a component of the telomere nucleoprotein complex. This protein is present at telomeres throughout the cell cycle and functions as an inhibitor of telomerase, acting in cis to limit the elongation of individual chromosome ends. The protein structure contains a C-terminal Myb motif, a dimerization domain near its N-terminus and an acidic N-terminus. Two transcripts of this gene are alternatively spliced products. [provided by RefSeq, Jul 2008]|
|NCBI GenInfo Identifier:||206729904|
|NCBI Gene ID:||7013|
|UniProt Secondary Accession:||P54274,Q15553, Q8NHT6, Q93029, A7XP29,|
|UniProt Related Accession:||P54274|
|NCBI Full Name:||Telomeric repeat-binding factor 1|
|NCBI Synonym Full Names:||telomeric repeat binding factor (NIMA-interacting) 1|
|NCBI Official Symbol:||TERF1|
|NCBI Official Synonym Symbols:||TRF; PIN2; TRF1; TRBF1; t-TRF1; hTRF1-AS|
|NCBI Protein Information:||telomeric repeat-binding factor 1; NIMA-interacting protein 2; telomeric protein Pin2/TRF1; TTAGGG repeat-binding factor 1|
|UniProt Protein Name:||Telomeric repeat-binding factor 1|
|UniProt Synonym Protein Names:||NIMA-interacting protein 2; TTAGGG repeat-binding factor 1; Telomeric protein Pin2/TRF1|
|Protein Family:||Telomeric repeat-binding factor|
|UniProt Gene Name:||TERF1|
|UniProt Entry Name:||TERF1_HUMAN|