PA5-46681

antibody from Invitrogen Antibodies

Targeting: FAM47E FLJ42946, LOC100129583

Provider product page for PA5-46681

Western blot

Other assay

Antibody data

Antibody Data
Antigen structure
References [1]
Comments [0]
Validations
Western blot [1]
Other assay [6]

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Product number: PA5-46681 - Provider product page
Provider: Invitrogen Antibodies
Product name: FAM47E Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: Peptide sequence: KLEDTWAYCQ DTRKGMKEPT KLLKKHSTQV YLGPSKKTSV SNAGQWLYEE
Concentration: 0.5 mg/mL

FAM47E protein structure - PA5-46681 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1. PRMT5 interacts with FAM47E. (A) Y2H assay was performed to study the interaction between PRMT5 with FAM47E. The pGBKT7-PRMT5 and PGADT7-FAM47E constructs were allowed to interact with each other or corresponding vector controls. The positive interaction was assessed by the investigating the expression of reporter genes. SD-Trp/-Leu denotes the synthetically defined medium which lacks tryptophan and leucine, SD-Trp/-Leu/+Aba/+X-alpha-Gal denotes synthetically defined medium which lacks tryptophan and leucine but contains Aureobasidin A and X-alpha-Gal and SD-Trp/-Leu/-His/-Ade/+Aba/+X-alpha-Gal denotes synthetically defined medium which lacks tryptophan, leucine, histidine, and adenine but contains Aureobasidin A and X-alpha-Gal. (B) HEK293 cells were co-transfected with Myc-PRMT5 and GFP or GFP-FAM47E constructs. Co-immunoprecipitation (Co-IP) was performed using GFP-Trap and the bound fractions were probed with Myc antibody. About 0.5% of the whole cell lysates which were used in Co-IP were probed with Myc antibody or GFP antibody. (C) HEK293 cells were co-transfected with HA-FAM47E and GFP or GFP-PRMT5 constructs. Co-IP was performed using GFP-Trap and the bound fractions were probed with HA antibody. About 1% of the whole cell lysates which were used in Co-IP were probed with HA antibody or GFP antibody. (D) Endogenous PRMT5 interacts with GFP tagged FAM47E. HEK293 cells were transfected with GFP or GFP-FAM47E constructs. Co-IP was performed using GFP-Trap and t

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2. FAM47E increases the stability of PRMT5 protein. (A) HEK293 cells were transfected with GFP vector or GFP-FAM47E construct. After 48 h of transfection, the cells were lysed, immunoblotted, and probed with PRMT5 antibody or GFP antibody or beta actin antibody (upper panel). The band intensities of PRMT5 and beta actin in the blots were quantified using ImageJ software and the relative ratios of PRMT5 signal to beta actin signal are plotted in the graph (lower panel). The values represent the mean of three independent experiments, with error bars representing SD. Statistical significance was assessed using two-tailed t test. * indicates P < 0.05. (B) HEK293 cells were transfected with control siRNA vector or FAM47E siRNA. After 48 h of transfection, the cells were lysed, immunoblotted, and probed with PRMT5 antibody or FAM47E antibody or beta actin antibody (upper panel). The band intensities of PRMT5 and beta actin in the blots were quantified using ImageJ software and the relative ratios of PRMT5 signal to beta actin signal are plotted in the graph (lower panel). The values represent the mean of three independent experiments, with error bars representing SD. Statistical significance was assessed using two-tailed t test. ** indicates P < 0.01. (C) HEK293 cells were transfected with GFP vector and GFP-FAM47E construct. After 40 h of transfection, the cells were treated with DMSO or MG-132 and incubated for 8 h. The cells were lysed, immunoblotted and probed with PRMT5

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3. FAM47E enhances the chromatin association of PRMT5. (A) HEK293 cells were transfected with, GFP vector or GFP-FAM47E construct or control siRNA or FAM47E siRNA. After 48 h of transfection, the cells were lysed, immunoblotted, and probed with GFP antibody or beta actin antibody (left panel) and FAM47E antibody or beta actin antibody (right panel). (B) HEK293 cells were transfected with GFP vector or GFP-FAM47E construct or control siRNA or FAM47E siRNA. After 48 h of transfection, the cells were lysed, the soluble fractions of the nuclei were isolated, and immunoblotting was performed using PRMT5 antibody or beta actin antibody (upper panels). The band intensities of PRMT5 and beta actin in the blots were quantified using ImageJ software and the relative ratios of PRMT5 signal to beta actin signal are plotted in the graph (lower panels). The values represent the mean of three independent experiments, with error bars representing standard deviations. Statistical significance was assessed using two-tailed t test. * indicates P < 0.05 and ** indicates P < 0.01. (C) HEK293 cells were transfected with GFP vector or GFP-FAM47E construct or control siRNA or FAM47E siRNA. After 48 h of transfection, the cells were lysed, the chromatin fractions were prepared by benzonase digestion and immunoblotting was performed using PRMT5 antibody or histone 3 antibody (upper panels). The band intensities of PRMT5 and histone 3 in the blots were quantified using ImageJ software and the re

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5. FAM47E regulates the expression of PRMT5 target genes. (A, B) HEK293 cells were transfected with GFP vector or GFP-FAM47E construct or control siRNA or FAM47E siRNA or co-transfected with GFP-FAM47E construct and control siRNA or PRMT5 siRNA. After 48 h of transfections, the whole cell lysate and total RNA were isolated from these cells. (A) The cell lysates were immunoblotted and probed with GFP antibody or PRMT5 antibody or FAM47E antibody or beta actin antibody. (B) The transcripts levels of the indicated PRMT5 target genes in these cells were quantified by using quantitative RT-PCR. The mRNA levels of indicated PRMT5 target genes were normalized to GAPDH expression and are presented relative to the control sample. The values represent the mean of three independent experiments, with error bars representing standard deviations. Statistical significance was assessed using two-tailed t test. * indicates P < 0.05, ** indicates P < 0.01, and n.s. indicates not significant. (C) The chromatin was prepared from HEK293 cells and the ChIP was performed using mouse IgG or PRMT5 antibody or rabbit IgG or FAM47E antibody. The association of PRMT5 and FAM47E with the promoters of PRMT5 target genes was investigated by analyzing the immunoprecipitated DNA using quantitative RT-PCR. Data are presented as fold enrichment relative to the control IgG binding and the values represent the mean of three independent experiments, with error bars representing standard deviations.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6. FAM47E increases cell proliferation and the clonogenic potential of HeLa cells through PRMT5. (A) HeLa cells were transfected with GFP vector or GFP-FAM47E construct or control siRNA or FAM47E siRNA or co-transfected with GFP-FAM47E construct and control siRNA or PRMT5 siRNA. After 48 h of transfection, the cells were lysed and probed with GFP antibody or PRMT5 antibody or FAM47E antibody or beta actin antibody. (B) HeLa cells were transfected with GFP vector or GFP-FAM47E construct or control siRNA or FAM47E siRNA or co-transfected with GFP-FAM47E construct and control siRNA or PRMT5 siRNA. The cells were counted after 48, 72 and 96 h of post-transfection ( Fig S12 ) and the doubling times were calculated. The values in the graph represent the mean of three independent experiments, with error bars representing SD. Statistical significance was assessed using two-tailed t test. ** indicates P < 0.01 and *** indicates P < 0.001. (C) HeLa cells were transfected with GFP vector or GFP-FAM47E construct or control siRNA or FAM47E siRNA or co-transfected with GFP-FAM47E construct and control siRNA or PRMT5 siRNA. After 48 h of transfections, MTT assay was carried out. The values in the graph represent the mean of three independent experiments, with error bars representing standard deviations. Statistical significance was assessed using two-tailed t test. ** indicates P < 0.01 and *** indicates P < 0.001. (D) HeLa cells were transfected with GFP vector or GFP-FAM47E constru

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure S9. FAM47E is localized both in the cytoplasm and nucleus. Representative image showing the sub-cellular localization of FAM47E in HEK293 cells. DAPI is shown in blue and the FAM47E staining is shown in red. Scale bar is shown on the image.