Anti-CTCF Antibody Picoband™

Product Name

Anti-CTCF Antibody Picoband™

View all CTCF Antibodies

SKU/Catalog Number

PB9493

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-CTCF Antibody Picoband™ catalog # PB9493. Tested in IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-CTCF Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9493)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human CTCF recombinant protein (Position: K521-R727). Human CTCF shares 94% and 93.1% amino acid (aa) sequence identity with mouse and rat CTCF, respectively.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PB9493 is reactive to CTCF in Human, Mouse, Rat

Applications

PB9493 is guaranteed for IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

140 kDa

Calculated molecular weight

82.785kDa

Background of CTCF

Transcriptional repressor CTCF also known as 11-zinc finger protein or CCCTC-binding factor is a transcription factor that in humans is encoded by the CTCF gene. This gene is a member of the BORIS + CTCF gene family and encodes a transcriptional regulator protein with 11 highly conserved zinc finger (ZF) domains. And this nuclear protein is able to use different combinations of the ZF domains to bind different DNA target sequences and proteins. Depending upon the context of the site, the protein can bind a histone acetyltransferase (HAT)-containing complex and function as a transcriptional activator or bind a histone deacetylase (HDAC)-containing complex and function as a transcriptional repressor. If the protein is bound to a transcriptional insulator element, it can block communication between enhancers and upstream promoters, thereby regulating imprinted expression. Mutations in this gene have been associated with invasive breast cancers, prostate cancers, and Wilms' tumors. Alternatively spliced transcript variants encoding different isoforms have been found for this gene.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of CTCF using anti-CTCF antibody (PB9493).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human 293T whole cell lysates,
Lane 2: human Caco-2 whole cell lysates,
Lane 3: rat C6 whole cell lysates,
Lane 4: mouse brain tissue lysates,
Lane 5: mouse RAW264.7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CTCF antigen affinity purified polyclonal antibody (Catalog # PB9493) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CTCF at approximately 140 kDa. The expected band size for CTCF is at 83 kDa.

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Figure 2. IHC analysis of CTCF using anti-CTCF antibody (PB9493).
CTCF was detected in a paraffin-embedded section of Mouse Intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-CTCF Antibody (PB9493) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of CTCF using anti-CTCF antibody (PB9493).
CTCF was detected in a paraffin-embedded section of Rat Intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-CTCF Antibody (PB9493) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 4. IHC analysis of CTCF using anti-CTCF antibody (PB9493).
CTCF was detected in a paraffin-embedded section of Human Lung Cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-CTCF Antibody (PB9493) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 5. IF analysis of CTCF using anti-CTCF antibody (PB9493).
CTCF was detected in an immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-CTCF Antibody (PB9493) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Gene/Protein Information For CTCF (Source: Uniprot.org, NCBI)

Alternative Names

11 zinc finger transcriptional repressor; 11-zinc finger protein; 11-Zinc Finger Protein2; CCCTC-binding factor (zinc finger protein); CCCTC-binding Factor; CTCF; CTCFL paralog; transcriptional repressor CTCF CTCF CFAP108, FAP108, MRD21 CCCTC-binding factor transcriptional repressor CTCF|11 zinc finger transcriptional repressor|11-zinc finger protein|CCCTC-binding factor (zinc finger protein)|CTCFL paralog

For more info on CTCF, check out the CTCF Infographic

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In this infographic, you will see the following information for CTCF: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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