PUF60 Rabbit Polyclonal Antibody
cat.: ER1915-62
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Tissue, FC
Clonality: Polyclonal
Form: Liquid
Storage condition: 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℃.
Storage buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: 60 KDa
Isotype: IgG
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.
Recommended Dilutions:
  WB
  IHC-P
  IF-tissue
  FC
1:500-2000
1:100-500
1:100-500
2ug/Test
Uniprot #: SwissProt: Q9UHX1 Human
Alternative names: 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 60 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
Images
ER1915-62_1.jpg Fig1: Sample:
Hela(Human) Cell Lysate at 30 ug
Primary: Anti-PUF60 (bs-19595R) 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_2.jpg Fig2: Sample:
Hela(Human) Cell Lysate at 30 ug
Primary: Anti-PUF60 (bs-19595R) 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.jpg Fig3: Sample:
LOVO(Human) Cell Lysate at 30 ug
Primary:Anti-PUF60 (bs-19595R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kD
Observed band size: 64 kD
ER1915-62_4.jpg Fig4: 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 (bs-19595R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
ER1915-62_5.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°C for 30min; Antibody incubation with (PUF60) Polyclonal Antibody, Unconjugated (bs-19595R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
ER1915-62_6.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°C for 30min; Antibody incubation with (PUF60) Polyclonal Antibody, Unconjugated (bs-19595R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
ER1915-62_7.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°C for 30min; Antibody incubation with (PUF60) Polyclonal Antibody, Unconjugated (bs-19595R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
ER1915-62_8.jpg Fig8: Blank control:Hela.
Primary Antibody (green line): Rabbit Anti-PUF60 antibody (bs-19595R)
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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.