Human TERF2 (Telomeric Repeat Binding Factor 2) ELISA Kit (HUES02822)
- SKU:
- HUES02822
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- Q15554
- Sensitivity:
- 0.09ng/mL
- Range:
- 0.16-10ng/mL
- ELISA Type:
- Sandwich
- Synonyms:
- TRBF2, TRF2
- Reactivity:
- Human
- Tested Sample Types:
- Serum, plasma and other biological fluids
Description
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 TERF2 in samples. No significant cross-reactivity or interference between Human TERF2 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 TERF2. 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 TERF2 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 TERF2, 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 TERF2. The concentration of Human TERF2 in samples can be calculated by comparing the OD of the samples to the standard curve.
UniProt Protein Function: | TRF2: Binds the telomeric double-stranded 5'-TTAGGG-3' repeat and plays a central role in telomere maintenance and protection against end-to-end fusion of chromosomes. In addition to its telomeric DNA-binding role, required to recruit a number of factors and enzymes required for telomere protection, including the shelterin complex, TERF2IP/RAP1 and DCLRE1B/Apollo. Component of the shelterin complex (telosome) that is involved in the regulation of telomere length and protection. Shelterin associates with arrays of double-stranded 5'-TTAGGG-3' repeats added by telomerase and protects chromosome ends; without its protective activity, telomeres are no longer hidden from the DNA damage surveillance and chromosome ends are inappropriately processed by DNA repair pathways. Together with DCLRE1B/Apollo, plays a key role in telomeric loop (T loop) formation by generating 3' single- stranded overhang at the leading end telomeres: T loops have been proposed to protect chromosome ends from degradation and repair. Required both to recruit DCLRE1B/Apollo to telomeres and activate the exonuclease activity of DCLRE1B/Apollo. Preferentially binds to positive supercoiled DNA. Together with DCLRE1B/Apollo, required to control the amount of DNA topoisomerase (TOP1, TOP2A and TOP2B) needed for telomere replication during fork passage and prevent aberrant telomere topology. Recruits TERF2IP/RAP1 to telomeres, thereby participating in to repressing homology- directed repair (HDR), which can affect telomere length. Homodimer. Component of the shelterin complex (telosome) composed of TERF1, TERF2, TINF2, TERF2IP/RAP1, ACD and POT1. Interacts with TERF2IP. Interacts with NBN. Interacts with SLX4/BTBD12. Interacts with DCLRE1B/Apollo and TERF2IP/RAP1; the interaction is direct. Ubiquitous. Highly expressed in spleen, thymus, prostate, uterus, testis, small intestine, colon and peripheral blood leukocytes. 2 isoforms of the human protein are produced by alternative splicing. |
UniProt Protein Details: | Protein type:DNA-binding; DNA repair, damage Chromosomal Location of Human Ortholog: 16q22. 1 Cellular Component: nucleoplasm; male germ cell nucleus; Golgi apparatus; chromosome, telomeric region; nuclear telomere cap complex; Mre11 complex; nuclear chromosome, telomeric region; cytoplasm; nucleus Molecular Function:protein C-terminus binding; protein binding; protein homodimerization activity; telomeric DNA binding; double-stranded telomeric DNA binding; chromatin binding Biological Process: negative regulation of telomere maintenance; telomere capping; negative regulation of telomere maintenance via semi-conservative replication; in utero embryonic development; telomeric loop formation; positive regulation of nitric-oxide synthase activity; positive regulation of telomere maintenance; telomere maintenance via telomerase; cell cycle; age-dependent telomere shortening; telomere maintenance; protection from non-homologous end joining at telomere |
NCBI Summary: | This gene encodes a telomere specific protein, TERF2, which is a component of the telomere nucleoprotein complex. This protein is present at telomeres in metaphase of the cell cycle, is a second negative regulator of telomere length and plays a key role in the protective activity of telomeres. While having similar telomere binding activity and domain organization, TERF2 differs from TERF1 in that its N terminus is basic rather than acidic. [provided by RefSeq, Jul 2008] |
UniProt Code: | Q15554 |
NCBI GenInfo Identifier: | 613504836 |
NCBI Gene ID: | 7014 |
NCBI Accession: | Q15554. 3 |
UniProt Related Accession: | Q15554 |
Molecular Weight: | 500 |
NCBI Full Name: | Telomeric repeat-binding factor 2 |
NCBI Synonym Full Names: | telomeric repeat binding factor 2 |
NCBI Official Symbol: | TERF2 |
NCBI Official Synonym Symbols: | TRF2; TRBF2 |
NCBI Protein Information: | telomeric repeat-binding factor 2; telomeric DNA-binding protein; TTAGGG repeat-binding factor 2; telomeric repeat binding protein 2 |
UniProt Protein Name: | Telomeric repeat-binding factor 2 |
UniProt Synonym Protein Names: | TTAGGG repeat-binding factor 2; Telomeric DNA-binding protein |
Protein Family: | Tricorn protease-interacting factor |
UniProt Gene Name: | TERF2 |
UniProt Entry Name: | TERF2_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.D | Average | Corrected |
10 | 2.353 2.387 | 2.37 | 2.281 |
5 | 1.513 1.555 | 1.534 | 1.445 |
2.5 | 0.871 0.831 | 0.851 | 0.762 |
1.25 | 0.458 0.496 | 0.477 | 0.388 |
0.63 | 0.264 0.252 | 0.258 | 0.169 |
0.32 | 0.194 0.166 | 0.18 | 0.091 |
0.16 | 0.137 0.137 | 0.137 | 0.048 |
0 | 0.082 0.096 | 0.089 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human TERF2 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human TERF2 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 (ng/mL) | 0.49 | 1.60 | 3.38 | 0.46 | 1.52 | 3.22 |
Standard deviation | 0.03 | 0.09 | 0.14 | 0.03 | 0.06 | 0.15 |
C V (%) | 6.12 | 5.63 | 4.14 | 6.52 | 3.95 | 4.66 |
Recovery
The recovery of Human TERF2 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) | 95-112 | 102 |
EDTA plasma (n=5) | 96-108 | 101 |
Cell culture media (n=5) | 94-105 | 100 |
Linearity
Samples were spiked with high concentrations of Human TERF2 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 (%) | 92-107 | 93-108 | 85-95 |
Average (%) | 98 | 101 | 90 | |
1:4 | Range (%) | 92-102 | 79-91 | 86-97 |
Average (%) | 97 | 86 | 92 | |
1:8 | Range (%) | 85-101 | 86-99 | 87-100 |
Average (%) | 92 | 92 | 93 | |
1:16 | Range (%) | 88-106 | 82-96 | 89-102 |
Average (%) | 96 | 88 | 94 |
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 |
- 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 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.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.