Antibodies, Proteins, Kits and Reagents for Life Science

294 835 ₸ Product size

100 µl 294 835 ₸ 10 µl 117 264 ₸

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Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18253] to CTCF - ChIP Grade
Suitable for: ChIP-sequencing, IHC-P, ChIP, ICC/IF, WB
Reacts with: Mouse, Rat, Human

Product name

Anti-CTCF antibody [EPR18253] - ChIP Grade
See all CTCF primary antibodies
Description

Rabbit monoclonal [EPR18253] to CTCF - ChIP Grade
Host species

Rabbit

Tested Applications & Species

Application	Species
ChIP	Human
ChIP-seq	Human
ICC/IF	Mouse Human
IHC-P	Mouse Rat Human
WB	Mouse Rat Human

See all applications and species data

Positive control
- WB: HeLa, LLC, RAW 264.7, HEK-293, HepG2, MCF7, C6, PC-12 and NIH/3T3 whole cell lysates; human fetal brain, fetal heart, fetal kidney and fetal spleen lysates; mouse brain, heart, kidney and spleen lysates; rat brain, heart, kidney and spleen lysates. IHC-P: Human endometrium, mouse liver and rat stomach tissues. ICC/IF: HeLa and NIH/3T3 cells. ChIP: Chromatin prepared from HeLa cells. ChIP-seq: Chromatin prepared from Hela cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18253
Isotype

IgG
Research areas

ChIP - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

Chromatin was prepared from HeLa (Human epithelial cells from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab188408 (blue), and 20µl of Anti Rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
ChIP-sequencing - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 30 µg of chromatin and 4 µg of Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408). ChIP DNA was sequenced on the Illumina NextSeq 500 to a depth of 30 million reads. ChIP-Seq validation performed by Active Motif, Carlsbad, CA.

Additional screenshots of mapped reads can be downloaded here.
Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

All lanes : Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408) at 1/5000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : LLC (Mouse lung carcinoma) whole cell lysate
Lane 3 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 4 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
Lane 5 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
Lane 6 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 83, 46 kDa

Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.
Observed MW: 140, 97, 80, 73, 70, 55 kDa.
The multiples bands observed in the WB have been reported in literature (PMID: 12878173, PMID: 8246978).
Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

All lanes : Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408) at 1/1000 dilution

Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 83, 46 kDa

Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.
Observed MW: 140, 130, 97, 80, 73, 70, 55 kDa.
The multiples bands observed in the WB have been reported in literature (PMID: 12878173, PMID: 8246978).
Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

All lanes : Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408) at 1/1000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human fetal spleen lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/100000 dilution

Predicted band size: 83, 46 kDa

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Observed MW: 140, 130, 97, 80, 73, 70, 55 kDa.
The multiples bands observed in the WB have been reported in literature (PMID: 12878173, PMID: 8246978).
Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

All lanes : Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408) at 1/1000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Mouse spleen lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 83, 46 kDa

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Observed MW: 140, 130, 97, 80, 73, 70, 55 kDa.
The multiples bands observed in the WB have been reported in literature (PMID: 12878173, PMID: 8246978).
Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

All lanes : Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408) at 1/1000 dilution

Lane 1 : Rat brain lysate
Lane 2 : Rat heart lysate
Lane 3 : Rat kidney lysate
Lane 4 : Rat spleen lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 83, 46 kDa

Exposure time: 2 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Observed MW 140, 130, 97, 80, 73, 70, 55 kDa.
The multiples bands observed in the WB have been reported in literature (PMID: 12878173, PMID: 8246978).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling CTCF with ab188408 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human endometrium is observed. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling CTCF with ab188408 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on hepatocytes of mouse liver is observed. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling CTCF with ab188408 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on the epithelium cells of rat stomach is observed. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling CTCF with ab188408 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab188408 at 1/2000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: ab7291 Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyronic fibroblast cells) cells labeling CTCF with ab188408 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) seconday antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab188408 at 1/2000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

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