重组Anti-CTCF抗体[EPR7314(B)] - ChIP Grade - BSA and Azide free (ab240035)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7314(B)] to CTCF - ChIP Grade – BSA and Azide free
- Suitable for: ChIP-sequencing, ChIP, Flow Cyt (Intra), IHC-P, ChIC/CUT&RUN-seq, ICC/IF, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-CTCF抗体[EPR7314(B)] - ChIP Grade - BSA and Azide free
参阅全部 CTCF 一抗 -
描述
兔单克隆抗体[EPR7314(B)] to CTCF - ChIP Grade – BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: ChIP-sequencing, ChIP, Flow Cyt (Intra), IHC-P, ChIC/CUT&RUN-seq, ICC/IF, WBmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab209492) -
阳性对照
- WB: HeLa and 293T cell lysates, Human colon tissue, Mouse brain tissue and rat heart tissue. IHC: Human breast carcinoma, mouse and rat kideny ICC/IF: HeLa cell lysates Flow Cyt (intra): 293T cell lysates ChIP-seq: HeLa cells. ChIC/CUT&RUN-Seq: HeLa cells.
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常规说明
ab240035 is the carrier-free version of ab128873.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
解离常数(KD)
KD = 1.74 x 10 -11 M Learn more about KD -
存储溶液
pH: 7.2
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR7314(B) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Anti-CTCF antibody [EPR7314(B)] - ChIP Grade (ab128873)
- Alexa Fluor® 488 Anti-CTCF antibody [EPR7314(B)] (ab203704)
- Alexa Fluor® 647 Anti-CTCF antibody [EPR7314(B)] (ab203705)
- PE Anti-CTCF antibody [EPR7314(B)] (ab305764)
- APC Anti-CTCF antibody [EPR7314(B)] (ab305765)
- HRP Anti-CTCF antibody [EPR7314(B)] (ab305766)
- Alexa Fluor® 594 Anti-CTCF antibody [EPR7314(B)] (ab310547)
- Alexa Fluor® 555 Anti-CTCF antibody [EPR7314(B)] (ab312077)
- Alexa Fluor® 568 Anti-CTCF antibody [EPR7314(B)] (ab312554)
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Compatible Secondaries
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Conjugation kits
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Immunizing Peptide (Blocking)
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Immunohistochemistry kits
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab240035于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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ChIP-sequencing |
Use 4 µg for 30 µg of chromatin.
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ChIP |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 140 kDa (predicted molecular weight: 83 kDa).Can be blocked with CTCF peptide (ab209492).
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说明 |
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ChIP-sequencing
Use 4 µg for 30 µg of chromatin. |
ChIP
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 140 kDa (predicted molecular weight: 83 kDa).Can be blocked with CTCF peptide (ab209492). |
靶标
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功能
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 to 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. 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. -
组织特异性
Ubiquitous. Absent in primary spermatocytes. -
序列相似性
Belongs to the CTCF zinc-finger protein family.
Contains 11 C2H2-type zinc fingers. -
结构域
The 11 zinc fingers are highly conserved among vertebrates, exhibiting almost identical amino acid sequences. Different subsets or combination of individual zinc fingers gives the ability to CTCF to recognize multiple DNA target sites. -
翻译后修饰
Sumoylated on Lys-74 and Lys-689; sumoylation of CTCF contributes to the repressive function of CTCF on the MYC P2 promoter. -
细胞定位
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. - Information by UniProt
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数据库链接
- Entrez Gene: 10664 Human
- Entrez Gene: 13018 Mouse
- Entrez Gene: 83726 Rat
- Omim: 604167 Human
- SwissProt: P49711 Human
- SwissProt: Q61164 Mouse
- SwissProt: Q9R1D1 Rat
- Unigene: 368367 Human
see all -
别名
- 11 zinc finger protein antibody
- 11 zinc finger transcriptional repressor antibody
- 11-zinc finger protein antibody
see all
图片
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This data was developed using the same antibody clone in a different buffer formulation (ab128873).
CUT&Tag-seq was performed using 200,000 Oli-neu (Oligodendrocyte progenitor) cells. Cells were permeabilized with 0.05% Digitonin and 0.01% NP-40 for 3 minutes. A 1:100 dilution of Recombinant Anti-CTCF antibody [EPR7314(B)] - ChIP Grade (ab128873) was used, along with a Guinea pig anti-rabbit Secondary. DNA was seg using Illumina NovaSeq S Prime to a depth of 24 million reads.
This image is courtesy of Dr Marek Bartosovic, Gonçalo Castelo-Branco Group, Karolinska Institutet.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells and 2 µg of ab128873 [EPR7314(B)]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The ChIP data was conducted on chromatin prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 HeLa cells and 4 µg of ab128873. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million readsAdditional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128873).
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Intracellular Flow Cytometry analysis showing 4% paraformaldehyde fixed 293T (human embryonic kidney) cells labelling CTCF with purified ab128873 at dilution of 1/40 followed by the secondary antibody; Alexa Fluor® 488 goat-anti-rabbit IgG at dilution of 1/500 (red line). A non-specific IgG antibody (rabbit monoclonal) was used as isotype control (black line). The blue line shows cells without incubation with primary antibody and secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128873).
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Immunocytochemistry/immunofluorescence staining of 4% paraformaldehyde fixed; 0.1% triton X 100 permeabilized HeLa (human cervix adenocarcinoma) cells with purified ab128873 at dilution of 1/500. The secondary antibody used was Alexa Fluor® 488; goat anti-rabbit IgG (ab150077) at a dilution of 1/1000. Nucleus was counter-stained with DAPI (blue). ab7291, a mouse anti-tubulin antibody (1/1000) was used to stain tubulin along with ab150120 (AlexaFluor®594 goat anti-mouse secondary, 1/1000) shown in the top right hand panel. The negative controls are shown in the bottom middle and right hand panels- for negative control 1 rabbit primary antibody and anti-mouse secondary antibody (ab150120) was used. For negative control 2 mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) was used.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128873).
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Immunohistochemical staining of paraffin-embedded rat kidney sections labelling CTCF with purified ab128873 at dilution of 1:1000. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128873).
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Immunohistochemical staining of paraffin-embedded mouse kidney sections labelling CTCF with purified ab128873 at dilution of 1:1000. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128873).
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Immunohistochemical staining of paraffin-embedded human breast carcinoma sections labelling CTCF with purified ab128873 at dilution of 1:1000. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128873).
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Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab128873 (red), and 20µl of protein A/G sepharose beads slurry (10µl of sepharose A beads + 10µl of sepharose G beads). 5μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128873).
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Overlay histogram showing Hela cells stained with un-purified ab128873 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab128873, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128873).
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Immunohistochemical analysis of paraffin embedded Human colon tissue labelling CTCF with un-purified ab128873 at 1/250 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128873).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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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 ab128873. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. ChIP-Seq validation performed with ChIP Kit Transcription Factors ChIP-Seq (ab270813).
Additional screenshots of mapped reads can be downloaded here. -
Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KDThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128873).
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (0)
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