Rabbit Anti-PUF60 antibody
60 kDa poly(U)-binding-splicing factor; FBP-interacting repressor; FIR; FLJ31379; FUSE-binding protein-interacting repressor; Poly U binding splicing factor 60KDa; Poly(U)-binding-splicing factor PUF60; poly-U binding splicing factor PUF60; PUF60; PUF60_H
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Product Name PUF60 Chinese Name SIAHBinding protein1抗体 Alias 60 kDa poly(U)-binding-splicing factor; FBP-interacting repressor; FIR; FLJ31379; FUSE-binding protein-interacting repressor; Poly U binding splicing factor 60KDa; Poly(U)-binding-splicing factor PUF60; poly-U binding splicing factor PUF60; PUF60; PUF60_HUMAN; pyrimidine tract binding splicing factor; Ro ribonucleoprotein-binding protein 1; Ro-binding protein 1; RoBP1; RoBPI; siah binding protein 1; Siah-binding protein 1; Siah-BP1; SIAHBP1. Research Area Tumour transcriptional regulatory factor Binding protein Epigenetics Immunogen Species Rabbit Clonality Polyclonal React Species Human, Mouse, Rat, Applications WB=1:500-2000 ELISA=1:5000-10000 IHC-P=1:100-500 IHC-F=1:100-500 Flow-Cyt=2ug/Test IF=1:100-500 （Paraffin sections need antigen repair）
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.Theoretical molecular weight 60kDa Cellular localization The nucleus Form Liquid Concentration 1mg/ml immunogen KLH conjugated synthetic peptide derived from human PUF60: 1-100/559 Lsotype IgG Purification affinity purified by Protein A Buffer Solution 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Storage Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. Attention This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. PubMed PubMed Product Detail This gene encodes a nucleic acid-binding protein that plays a role in a variety of nuclear processes, including pre-mRNA splicing and transcriptional regulation. The encoded protein forms a complex with the far upstream DNA element (FUSE) and FUSE-binding protein at the myelocytomatosis oncogene (MYC) promoter. This complex represses MYC transcription through the core-TFIIH basal transcription factor. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene. [provided by RefSeq, Aug 2012]
Function:
DNA- and RNA-binding protein, involved in several nuclear processes such as pre-mRNA splicing, apoptosis and transcription regulation. In association with FUBP1 regulates MYC transcription at the P2 promoter through the core-TFIIH basal transcription factor. Acts as a transcriptional repressor through the core-TFIIH basal transcription factor. Represses FUBP1-induced transcriptional activation but not basal transcription. Decreases ERCC3 helicase activity. Does not repress TFIIH-mediated transcription in xeroderma pigmentosum complementation group B (XPB) cells. Is also involved in pre-mRNA splicing. Promotes splicing of an intron with weak 3'-splice site and pyrimidine tract in a cooperative manner with U2AF2. Involved in apoptosis induction when overexpressed in HeLa cells. Isoform 6 failed to repress MYC transcription and inhibited FIR-induced apoptosis in colorectal cancer. Isoform 6 may contribute to tumor progression by enabling increased MYC expression and greater resistance to apoptosis in tumors than in normal cells. Modulates alternative splicing of several mRNAs. Binds to relaxed DNA of active promoter regions. Binds to the pyrimidine tract and 3'-splice site regions of pre-mRNA; binding is enhanced in presence of U2AF2. Binds to Y5 RNA in association with TROVE2. Binds to poly(U) RNA.
Subcellular Location:
Nucleus. Colocalizes partially with TROVE2.
Tissue Specificity:
Isoform 2 is expressed in colonic epithelium and colorectal epithelium cancer (at protein level). Isoform 6 is expressed in colorectal epithelial cancer but below detection level in colonic epithelium. Expressed in heart, brain, placenta, lung, liver, skeletal muscle, kidney, pancreas, spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes.
Similarity:
Belongs to the RRM half pint family.
Contains 3 RRM (RNA recognition motif) domains.
SWISS:
Q9UHX1
Gene ID:
22627
Database links:Entrez Gene: 22827 Human
Entrez Gene: 67959 Mouse
Omim: 604819 Human
SwissProt: Q9UHX1 Human
SwissProt: Q3UEB3 Mouse
Unigene: 521924 Human
Unigene: 29965 Mouse
Unigene: 96174 Rat
Product Picture 
Sample:
LOVO(Human) Cell Lysate at 30 ug
Primary:Anti-PUF60 (SL19595R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kD
Observed band size: 64 kD
Sample:
Hela(Human) Cell Lysate at 30 ug
Primary: Anti-PUF60 (SL19595R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kD
Observed band size: 60 kD
Sample:
Hela(Human) Cell Lysate at 30 ug
Primary: Anti-PUF60 (SL19595R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kD
Observed band size: 63 kD
Paraformaldehyde-fixed, paraffin embedded (Mouse colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PUF60) Polyclonal Antibody, Unconjugated (SL19595R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PUF60) Polyclonal Antibody, Unconjugated (SL19595R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PUF60) Polyclonal Antibody, Unconjugated (SL19595R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PUF60) Polyclonal Antibody, Unconjugated (SL19595R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Blank control:Hela.
Primary Antibody (green line): Rabbit Anti-PUF60 antibody (SL19595R)
Dilution: 2ug/Test;
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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