Human TERF1 (Telomeric repeat-binding factor 1) ELISA Kit (HUFI06147)
- SKU:
- HUFI06147
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P54274
- Sensitivity:
- 0.094ng/ml
- Range:
- 0.156-10ng/ml
- ELISA Type:
- Sandwich
- Synonyms:
- hTRF1 AS, NIMA interacting protein 2, PIN2, t TRF1, Telomeric protein Pin2, TRF1, TERF1, TRBF1, TRF, TRF1, TTAGGG repeat binding factor 1
- Reactivity:
- Human
Description
Human TERF1 (Telomeric repeat-binding factor 1) ELISA
TERF1 (Telomeric Repeat Binding Factor 1) is a protein that is essential for the replication of chromosomes. It binds to telomeric DNA and recruits other proteins necessary for replication. TERF1 has been shown to be essential for cell viability, and genetic studies have shown that disruption of TERF1 results in embryonic lethality. Diseases associated with TERF1 include Townes-Brocks Syndrome and Cherubism. Gene Ontology (GO) annotations related to TERF1 include protein homodimerization activity and chromatin binding. An important paralog of TERF1 is TERF2.
| Product Name: | Human TERF1 (Telomeric repeat-binding factor 1) ELISA Kit |
| Product Code: | HUFI06147 |
| Size: | 96 Assays |
| Alias: | hTRF1 AS ELISA Kit, NIMA interacting protein 2 ELISA Kit, PIN2 ELISA Kit, t TRF1 ELISA Kit, Telomeric protein Pin2 ELISA Kit, TRF1 ELISA Kit, TERF1 ELISA Kit, TRBF1 ELISA Kit, TRF ELISA Kit, TRF1 ELISA Kit, TTAGGG repeat binding factor 1 ELISA Kit |
| Detection method: | Sandwich ELISA, Double Antibody |
| Application: | This immunoassay kit allows for the in vitro quantitative determination of Human TERF1 (Telomeric repeat-binding factor 1) concentrations in serum plasma and other biological fluids. |
| Sensitivity: | < 0.094ng/ml |
| Range: | 0.156-10ng/ml |
| Storage: | 4°C for 6 months |
| Note: | For Research Use Only |
| Recovery: | Matrices listed below were spiked with certain level of Human TERF1 (Telomeric repeat-binding factor 1) and the recovery rates were calculated by comparing the measured value to the expected amount of Human TERF1 (Telomeric repeat-binding factor 1) in samples. Enquire for more information. |
| Linearity: | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human TERF1 (Telomeric repeat-binding factor 1) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Enquire for more information. |
| CV(%): | Intra-Assay: CV<8% Inter-Assay: CV<10% |
| Component | Quantity | Storage |
| ELISA Microplate (Dismountable) | 8×12 strips | 4°C for 6 months |
| Lyophilized Standard | 2 | 4°C/-20°C |
| Sample/Standard Dilution Buffer | 20ml | 4°C |
| Biotin-labeled Antibody(Concentrated) | 120ul | 4°C (Protect from light) |
| Antibody Dilution Buffer | 10ml | 4°C |
| HRP-Streptavidin Conjugate(SABC) | 120ul | 4°C (Protect from light) |
| SABC Dilution Buffer | 10ml | 4°C |
| TMB Substrate | 10ml | 4°C (Protect from light) |
| Stop Solution | 10ml | 4°C |
| Wash Buffer(25X) | 30ml | 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
| 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: | Protein type:DNA-binding 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] |
| UniProt Code: | P54274 |
| NCBI GenInfo Identifier: | 206729904 |
| NCBI Gene ID: | 7013 |
| NCBI Accession: | P54274.3 |
| UniProt Secondary Accession: | P54274,Q15553, Q8NHT6, Q93029, A7XP29, |
| UniProt Related Accession: | P54274 |
| Molecular Weight: | 439 |
| 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 |
*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37 °C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.
| Step | Protocol |
| 1. | Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells! |
| 2. | Aliquot 0.1ml standard solutions into the standard wells. |
| 3. | Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well. |
| 4. | Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells. |
| 5. | Seal the plate with a cover and incubate at 37 °C for 90 min. |
| 6. | Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2. |
| 7. | Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall. |
| 8. | Seal the plate with a cover and incubate at 37 °C for 60 min. |
| 9. | Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash. |
| 10. | Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37 °C for 30 min. |
| 11. | Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min. |
| 12. | Add 90 µL of TMB substrate into each well, cover the plate and incubate at 37 °C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color. |
| 13. | Add 50 µL of Stop solution into each well and mix thoroughly. The color changes into yellow immediately. |
| 14. | Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution. |
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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