Anti-PUF60 antibody
ER1915-62
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P
Molecular Wt: 60 KDa
Description:
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.
Immunogen:
KLH conjugated synthetic peptide derived from human PUF60 1-100/559
Positive control:
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.
Subcellular location:
Nucleus. Colocalizes partially with TROVE2.
Database links:
SwissProt: Q9UHX1 Human
Recommended Dilutions:
  

WB1:500-2000
IHC-P1:400-800
Storage Buffer:
0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
Storage Instruction:
Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20℃. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4℃.
Purity:
affinity purified by Protein A
Images
ER1915-62_1_small.jpg
Fig1: 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℃ for 30min; Antibody incubation with (PUF60) Polyclonal Antibody, Unconjugated (ER1915-62) at 1:400 overnight at 4℃, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
ER1915-62_2_small.jpg
Fig2: Sample:
Hela(Human) Cell Lysate at 30 ug
Primary: Anti-PUF60 (ER1915-62) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kD
Observed band size: 60 kD
ER1915-62_3_small.jpg
Fig3: Sample:
Hela(Human) Cell Lysate at 30 ug
Primary: Anti-PUF60 (ER1915-62) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kD
Observed band size: 63 kD
ER1915-62_4_small.jpg
Fig4: Sample:
LOVO(Human) Cell Lysate at 30 ug
Primary:Anti-PUF60 (ER1915-62) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kD
Observed band size: 64 kD
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ER1915-62_5_small.jpg
Fig5: 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℃ for 30min; Antibody incubation with (PUF60) Polyclonal Antibody, Unconjugated (ER1915-62) at 1:400 overnight at 4℃, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
ER1915-62_6_small.jpg
Fig6: 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℃ for 30min; Antibody incubation with (PUF60) Polyclonal Antibody, Unconjugated (ER1915-62) at 1:400 overnight at 4℃, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
ER1915-62_7_small.jpg
Fig7: 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℃ for 30min; Antibody incubation with (PUF60) Polyclonal Antibody, Unconjugated (ER1915-62) at 1:400 overnight at 4℃, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.
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