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Human CTCFL ELISA Kit

SKU:
HUFI01890
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q8NI51
Sensitivity:
0.188ng/ml
Range:
0.313-20ng/ml
ELISA Type:
Competitive
Synonyms:
CTCFL, Zinc finger protein CTCF-T, Cancer, testis antigen 27, CT27, CTCF-like protein, CTCF paralog, Brother of the regulator of imprinted sites, CCCTC-binding factor
Reactivity:
Human
Research Area:
Cell Cycle
€599
Frequently bought together:

Description

Human CTCFL ELISA Kit

The Human CTCFL ELISA Kit is specifically designed for the accurate detection of CTCFL (CTCF-like) levels in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, providing researchers with reliable and reproducible results for a variety of research applications.CTCFL is a key protein that plays a critical role in gene regulation and epigenetic processes. It is known to be involved in various biological functions such as chromatin organization, genomic imprinting, and embryonic development.

Aberrant expression of CTCFL has been linked to several diseases including cancer, developmental disorders, and infertility, making it a valuable biomarker for understanding disease mechanisms and exploring potential therapeutic interventions.With its advanced technology and user-friendly protocol, the Human CTCFL ELISA Kit is an essential tool for researchers studying CTCFL biology and its implications in health and disease. Get accurate and reliable results with this innovative kit from Assay Genie.

Product Name:Human CTCFL ELISA Kit
Product Code:HUFI01890
Size:96 Assays
Alias:CTCFL, Zinc finger protein CTCF-T, Cancer, testis antigen 27, CT27, CTCF-like protein, CTCF paralog, Brother of the regulator of imprinted sites, CCCTC-binding factor
Detection method:Competitive ELISA, Coated with Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human CTCFL concentrations in serum plasma and other biological fluids.
Sensitivity:0.188ng/ml
Range:0.313-20ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human CTCFL and the recovery rates were calculated by comparing the measured value to the expected amount of Human CTCFL in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)91-9994
EDTA plasma(n=5)86-9692
UFH plasma(n=5)95-10599
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human CTCFL and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)90-102%85-97%91-105%
EDTA plasma(n=5)82-88%84-99%85-100%
UFH plasma(n=5)82-100%83-100%81-100%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate(Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)60ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipettetips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotQ8NI51
UniProt Protein Function:BORIS: Testis-specific DNA binding protein responsible for insulator function, nuclear architecture and transcriptional control, which probably acts by recruiting epigenetic chromatin modifiers. Plays a key role in gene imprinting in male germline, by participating in the establishment of differential methylation at the IGF2/H19 imprinted control region (ICR). Directly binds the unmethylated H19 ICR and recruits the PRMT7 methyltransferase, leading to methylate histone H4 'Arg-3' to form H4R3sme2. This probably leads to recruit de novo DNA methyltransferases at these sites. Seems to act as tumor suppressor. In association with DNMT1 and DNMT3B, involved in activation of BAG1 gene expression by binding to its promoter. Required for dimethylation of H3 lysine 4 (H3K4me2) of MYC and BRCA1 promoters. Interacts with histones, PRMT7 and SETD1A. Interacts (via N-terminus) with BAG6/BAT3. Testis specific. Specifically expressed in primary spermatocytes. Belongs to the CTCF zinc-finger protein family.
UniProt Protein Details:Protein type:DNA-binding; Cancer Testis Antigen (CTA); C2H2-type zinc finger protein

Chromosomal Location of Human Ortholog: 20q13.31

Cellular Component: cytoplasm; nucleus

Molecular Function:protein binding; DNA binding; histone binding; sequence-specific DNA binding; metal ion binding

Biological Process: histone methylation; genetic imprinting; transcription, DNA-dependent; DNA methylation during gametogenesis; positive regulation of transcription, DNA-dependent; regulation of histone H3-K4 methylation; positive regulation of transcription from RNA polymerase II promoter; cell cycle

UniProt Code:Q8NI51
NCBI GenInfo Identifier:313104098
NCBI Gene ID:
NCBI Accession:Q8NI51.2
Molecular Weight:
NCBI Full Name:Transcriptional repressor CTCFL
UniProt Protein Name:Transcriptional repressor CTCFL
UniProt Synonym Protein Names:Brother of the regulator of imprinted sites; CCCTC-binding factor; CTCF paralog; CTCF-like protein; Cancer/testis antigen 27; CT27; Zinc finger protein CTCF-T
Protein Family:Transcriptional repressor
UniProt Gene Name:CTCFL  
UniProt Entry Name:CTCFL_HUMAN

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Equilibrate the TMB substrate for at least 30 min at 37°C beforeuse. When diluting samples and reagents, they must be mixed completely andevenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coatedplate respectively, and then, record their positions. It isrecommended to measure each standard and sample in duplicate. Washplate 2 times before adding standard, sample and control (zero) wells!
2.Add Sample and Biotin-detection antibody: Add 50µL of Standard, Blank or Sample per well. The blankwell is added with Sample Dilution Buffer. Immediately add 50 µL of biotin-labelled antibody workingsolution to each well. Cover with the plate sealer provided. Gently tap the plate to ensure thoroughmixing. Incubate for 45 minutes at 37°C. (Solutions are added to the bottom of micro-ELISA platewell, avoid touching plate walls and foaming).
3.Wash: Aspirate each well and wash, repeating the process three timesWash by filling each well with Wash Buffer (approximately 350µL)using a squirt bottle, multi-channel pipette, manifold dispenser orautomated washer. Complete removal of liquid at each step is essentialto good performance. After the last wash, remove any remaining WashBuffer by aspirating or decanting. Invert the plate and pat it againstthick clean absorbent paper.
4.HRP-Streptavidin Conjugate(SABC): Add 100µL of SABC workingsolution to each well. Cover with a new Plate sealer. Incubate for30minutes at 37°C.
5.Wash: Repeat the aspiration/wash process for five times.
6.TMB Substrate: Add 90µL of TMB Substrate to each well. Coverwith a new Plate sealer. Incubate for about 10-20 minutes at 37°C.Protect from light. The reaction time can be shortened or extendedaccording to the actual color change, but not more than 30minutes.When apparent gradient appeared in standard wells, you can terminatethe reaction.
7.Stop: Add 50µL of Stop Solution to each well. Color turn toyellow immediately. The adding order of stop solution should be as thesame as the substrate solution.
8.OD Measurement: Determine the optical density (OD Value) of each wellat once, using a microplate reader set to 450 nm. You should open themicroplate reader ahead, preheat the instrument, and set the testing parameters.

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 TypeProtocol
SerumIf 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.

PlasmaCollect 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 FluidCollect 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 supernatantCollect 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 lysatesSolubilize 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 homogenatesThe 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 lysatesRinse 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 MilkCollect 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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