PA5-88115

antibody from Invitrogen Antibodies

Targeting: CTCF CFAP108, FAP108

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-88115

Provider

Invitrogen Antibodies

Product name

CTCF Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Immunogen sequence: MEGDAVEAIV EESETFIKGK ERKTYQRRRE GGQEEDACHL PQNQTDGGEV VQDVNSSVQM VMMEQLDPTL LQMKTEVMEG TVAPEAEAAV DDTQIITLQV VNMEEQPINI GELQLVQVPV PVTVPVATTS VEELQGAYEN EVSKEGLAES EPMICHTLPL PEGFQVVKVG ANGEVETLEQ GELPPQEDPS WQKDPDYQPP AKKTKKTKKS KLRYTEEGKD VDVSVYDFEE EQQEGLLSEV NAEKVVGNMK PPKPTKIKKK; Positive Samples: 293T, Jurkat; Cellular Location: Chromosome, Nucleus, centromere, nucleoplasm

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.63 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

CTCF knockout in zebrafish induces alterations in regulatory landscapes and developmental gene expression.

Franke M, De la Calle-Mustienes E, Neto A, Almuedo-Castillo M, Irastorza-Azcarate I, Acemel RD, Tena JJ, Santos-Pereira JM, Gómez-Skarmeta JL
Nature communications 2021 Sep 13;12(1):5415

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry-Immunofluorescence analysis of CTCF was performed in U2OS cells using CTCF Polyclonal Antibody (Product # PA5-88115).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry-Immunofluorescence analysis of CTCF was performed in Hela cells using CTCF Polyclonal Antibody (Product # PA5-88115).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of CTCF was performed using 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with CTCF Polyclonal Antibody (Product # PA5-88115) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin immunoprecipitation analysis of CTCF in HCT116 using CTCF Polyclonal Antibody (Product # PA5-88115) and rabbit IgG. The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin Immunoprecipitation (ChIP) of CTCF protein was performed using CTCF Polyclonal Antibody (Product # PA5-88115, 5 µg) on sheared chromatin from MCF7 cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to H19-ICR, CRY1, PFKFB3, GLI2 (5'), AXL (3') (Active), EXON4/5 OF H19 and SAT2 satellite repeats (Inactive). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation analysis of CTCF was performed in 200 µg extracts of HeLa cells using CTCF Polyclonal Antibody (Product # PA5-88115). Western blot was performed from the immunoprecipitate using CTCF Polyclonal Antibody.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 1 Knockout of ctcf in zebrafish embryos disrupts chromatin structure. a Pictures of wild-type (WT), ctcf +/- and ctcf -/- zebrafish embryos at 48 h post fertilization (hpf) showing homozygous mutant phenotypes, including the reduced size of head and eyes, heart edema, and defective pigmentation (arrow heads). Scale bars represent 250 mum. b Whole-mount embryo immunofluorescence of CTCF (red) and dapi (blue) in WT, ctcf +/- and ctcf -/- zebrafish embryos at the stages of 1000 cells (1 K cells), 30% of epiboly (30% epib. ), 80% of epiboly (80% epib. ), 18 somites (18 som.) and 24 hpf showing the maternal contribution of CTCF protein. Relative quantification of CTCF/dapi signal with average values +- standard error is shown. Statistical significance was measured using a two-sided Student's t test. The number of embryos for WT, ctcf +/- and ctcf -/- used for quantification are as follows: 1 K cells ( n = 3, n = 12, n = 3); 30% epib. ( n = 4, n = 8, n = 5); 80% epib. ( n = 3, n = 10, n = 5); 18 som. ( n = 5, n = 5, n = 7) and 24 hpf ( n = 5, n = 17, n = 5). Source data are provided as a Source Data file. c HiC normalized contact maps at 10-kb resolution from WT and ctcf -/- zebrafish embryos at 24 and 48 hpf. A 3-Mb genomic region in chr11 is plotted, aligned with the insulation scores and the called topologically associating domain (TAD) boundaries. d Average insulation score profiles of WT and ctcf -/- zebrafish embryos at 24 and 48 hpf around the TAD boundaries called in t