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Mouse Transcriptional repressor CTCF (Ctcf) ELISA Kit (MOEB2113)

SKU:
MOEB2113
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q61164
Range:
78-5000 pg/mL
ELISA Type:
Sandwich
Synonyms:
Ctcf, Transcriptional repressor CTCF, CTCFL paralog, 11-zinc finger protein, CCCTC-binding factor
Reactivity:
Mouse
€649
Frequently bought together:

Description

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Mouse Transcriptional repressor CTCF (Ctcf) ELISA Kit

The Mouse Transcriptional Repressor CTCF (CTCF) ELISA Kit is a powerful tool designed for the accurate quantification of CTCF levels in mouse serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit delivers reliable and reproducible results, making it perfect for a wide range of research applications.CTCF is a critical transcriptional repressor involved in gene regulation and chromatin organization. It plays a crucial role in various biological processes such as development, differentiation, and disease progression.

By measuring CTCF levels, researchers can gain valuable insights into gene expression regulation and potentially identify new therapeutic targets for diseases like cancer, autoimmune disorders, and developmental abnormalities.Overall, the Mouse Transcriptional Repressor CTCF (CTCF) ELISA Kit is an essential tool for researchers looking to study the intricate mechanisms of gene regulation and chromatin dynamics in mouse models.

Product Name:Mouse Transcriptional repressor CTCF (Ctcf) ELISA Kit
SKU:MOEB2113
Size:96T
Target:Mouse Transcriptional repressor CTCF (Ctcf)
Synonyms:11-zinc finger protein, CCCTC-binding factor, CTCFL paralog
Assay Type:Competitive
Detection Method:ELISA
Reactivity:Mouse
Detection Range:78-5000pg/mL
Sensitivity:39.3pg/mL
Intra CV:7.2%
Inter CV:9.4%
Linearity:
Sample1:21:41:81:16
Serum(N=5)96-105%109-118%82-95%107-116%
EDTA Plasma(N=5)109-118%116-126%108-116%89-98%
Heparin Plasma(N=5)91-102%100-110%101-113%103-112%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum10397-109
Plasma10599-111
Function:Chromatin binding factor that binds to DNA sequence specific sites. Involved in transcriptional regulation by binding to chromatin insulators and preventing interaction between promoter and nearby enhancers and silencers. Acts as transcriptional repressor binding to promoters of vertebrate MYC gene and BAG1 gene. Also binds to the PLK and PIM1 promoters. Acts as a transcriptional activator of APP. Regulates APOA1/C3/A4/A5 gene cluster and controls MHC class II gene expression. Plays an essential role in oocyte and preimplantation embryo development by activating or repressing transcription. Seems to act as tumor suppressor. Plays a critical role in the epigenetic regulation. Participates in the allele-specific gene expression at the imprinted IGF2/H19 gene locus. On the maternal allele, binding within the H19 imprinting control region (ICR) mediates maternally inherited higher-order chromatin conformation to restrict enhancer access to IGF2. Plays a critical role in gene silencing over considerable distances in the genome. Preferentially interacts with unmethylated DNA, preventing spreading of CpG methylation and maintaining methylation-free zones. Inversely, binding to target sites is prevented by CpG methylation. Plays a important role in chromatin remodeling. Can dimerize when it is bound to different DNA sequences, mediating long-range chromatin looping (By similarity). Mediates interchromosomal association between IGF2/H19 and WSB1/NF1 and may direct distant DNA segments to a common transcription factory. Causes local loss of histone acetylation and gain of histone methylation in the beta-globin locus, without affecting transcription. When bound to chromatin, it provides an anchor point for nucleosomes positioning. Seems to be essential for homologous X-chromosome pairing. May participate with Tsix in establishing a regulatable epigenetic switch for X chromosome inactivation. May play a role in preventing the propagation of stable methylation at the escape genes from X-inactivation. Involved in sister chromatid cohesion. Associates with both centromeres and chromosomal arms during metaphase and required for cohesin localization to CTCF sites. Regulates asynchronous replication of IGF2/H19. Plays a role in the recruitment of CENPE to the pericentromeric/centromeric regions of the chromosome during mitosis.
Uniprot:Q61164
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant mouse Transcriptional repressor CTCF
Sub Unit:Interacts with CHD8 (PubMed:16949368). Interacts with LLPH (PubMed:26961175). Interacts with CENPE.
Research Area:Epigenetics
Subcellular Location:Nucleus Nucleoplasm Chromosome Chromosome Centromere May translocate to the nucleolus upon cell differentiation. Associates with both centromeres and chromosomal arms during metaphase. Associates with the H19 ICR in mitotic chromosomes. May be preferentially excluded from heterochromatin during interphase.
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:CTCF: a widely expressed, sequence-specific DNA-binding transcriptional regulator, insulator, and organizer of higher-order chromatin structure. Acts as a tumor suppressor. Contains 11 C2H2-type zinc fingers. Involved in promoter activation or repression, hormone-responsive gene silencing, methylation-dependent chromatin insulation, and genomic imprinting. Mediates pairing between X chromosomes and interactions between distant regulatory elements. Binds to promoters of c-myc, PLK, PIM1 and APP. A critical regulator of cell-cycle arrest and death after B cell receptor signaling in immature B cells. CTCF, together with YY1 and Tsix, form a regulated epigenetic switch for X-inactivation.
UniProt Protein Details:

Protein type:C2H2-type zinc finger protein; Transcription factor; Tumor suppressor

Chromosomal Location of Human Ortholog: 8|8 D3

Cellular Component: chromosome, pericentric region; condensed chromosome; nucleolus; nucleoplasm; nucleus

Molecular Function:chromatin binding; chromatin insulator sequence binding; DNA binding; protein binding; sequence-specific DNA binding; transcription factor activity

Biological Process: DNA methylation; dosage compensation, by inactivation of X chromosome; genetic imprinting; maintenance of DNA methylation; negative regulation of transcription from RNA polymerase II promoter; negative regulation of transcription, DNA-dependent; nucleosome positioning; positive regulation of transcription, DNA-dependent; regulation of gene expression, epigenetic; regulation of histone acetylation; regulation of histone methylation; regulation of molecular function, epigenetic; regulation of transcription, DNA-dependent

UniProt Code:Q61164
NCBI GenInfo Identifier:31044459
NCBI Gene ID:13018
NCBI Accession:NP_851839.1
UniProt Related Accession:Q61164
Molecular Weight:83,745 Da
NCBI Full Name:transcriptional repressor CTCF
NCBI Synonym Full Names:CCCTC-binding factor
NCBI Official Symbol:Ctcf
NCBI Official Synonym Symbols:AW108038
NCBI Protein Information:transcriptional repressor CTCF
UniProt Protein Name:Transcriptional repressor CTCF
UniProt Synonym Protein Names:11-zinc finger protein; CCCTC-binding factor; CTCFL paralog
Protein Family:Transcriptional repressor
UniProt Gene Name:Ctcf
Component Quantity (96 Assays) Storage
ELISA Microplate (Dismountable) 8×12 strips -20°C
Lyophilized Standard 2 -20°C
Sample Diluent 20ml -20°C
Assay Diluent A 10mL -20°C
Assay Diluent B 10mL -20°C
Detection Reagent A 120µL -20°C
Detection Reagent B 120µL -20°C
Wash Buffer 30mL 4°C
Substrate 10mL 4°C
Stop Solution 10mL 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

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

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.

Step
1. Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2. Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3. Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5. Repeat the wash process for five times as conducted in step 3.
6. Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7. Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8. Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9. After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

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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