Invitrogen

PAF49 Polyclonal Antibody

Product Image Gallery

PAF49 Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of PAF49 showing staining in the nucleolus of A431 cells. A431 cells were fixed in 4% paraformaldehyde at RT for 2 sec and stained using a PAF49 polyclonal antibody (Product # PA5-27632) diluted at 1:500. Blue: Hoechst 33342 staining.

PAF49 Antibody in Western Blot (WB) — Western blot analysis of PAF49 using Whole cell extracts (30 µg). Samples were loaded onto a 10% SDS-PAGE gel and probed with a PAF49 polyclonal antibody (Product # PA5-27632) at a dilution of 1:1000.

PAF49 Antibody in Immunoprecipitation (IP) — Immunoprecipitation of CD3EAP was performed in K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 to 10 µL of polyclonal CD3EAP antibody (Product # PA5-27632) rotating 60 min at RT. The immune complexes were captured on 625 µg of anti-rabbit coated Dynabeads (Product # 11204D) and washed extensively. They were then eluted and analyzed using the Simple Western system using the same antibody as used in immunoprecipitation at a dilution of 1:25, followed by a 1:100 dilution of secondary antibody. Lane 1 is the input, lane 2 no antibody IP and lane 3 is the target specific IP. Data courtesy of the Yeo lab as part of the ENCODE project.

PAF49 Antibody in ChIP assay (ChIP) — Chromatin immunoprecipitation analysis of PAF49 was performed using cross-linked chromatin from 1x10^6 HCT116 colon carcinoma cells treated with serum for 0 and 30 minutes. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0 µL/100 µL well volume of a PAF49 polyclonal antibody (Product # PA5-27632). Chromatin aliquots from ~1x10^5 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µL of eluted DNA in 2 µL SYBR real-time PCR reactions containing primers to amplify exon-1 of the RNA5-8S5 gene or -20 kb upstream of the LAMC1 gene. PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. A schematic representations of the RNA5-8S5 and LAMC1 loci are shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions), the zigzag line represents an intron, and the straight line represents upstream sequence. Regions amplified by RNA5-8S5 and LAMC1 primers are represented by black bars. Data courtesy of the Innovators Program.

Product Details

Tested Applications

Dilution

ChIP assay (ChIP)

Assay-Dependent

Immunocytochemistry (ICC)

1:100-1:1000

Immunofluorescence (IF)

1:100-1:1000

Western Blot (WB)

1:500-1:3000

RNA Immunoprecipitation (RIP)

5-10 µL per 500 µg lysate

Product Specifications

Species Reactivity

Human

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Recombinant fragment corresponding to a region within amino acids 1 and 268 of Human PAF49

Conjugate

Unconjugated

Form

Liquid

Concentration

0.55 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS, pH 7, with 20% glycerol

Contains

0.025% ProClin 300

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2545108

Product Specific Information

PA5-27632 targets PAF49 in IF, IHC (P), ChIP and WB applications and shows reactivity with Human samples.

The PA5-27632 immunogen is recombinant fragment corresponding to a region within amino acids 1 and 268 of Human PAF49.

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial. For short-term storage (1-2 weeks), product can be stored at 4°C. For long-term storage, aliquot and store product at -20° C or below, avioiding multiple freeze-thaw cycles.

Target Information

DNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. Component of RNA polymerase I which synthesizes ribosomal RNA precursors. Isoform 1 is involved in UBTF-activated transcription, presumably at a step following PIC formation. Isoform 2 has been described as a component of preformed T-cell receptor (TCR) complex.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: A34.5; antis; antisense to ERCC 1; Antisense to ERCC-1 protein; ASE-1; CAST; CD3 epsilon associated protein; CD3-epsilon-associated protein; CD3e antigen; CD3e antigen, epsilon polypeptide associated protein; CD3e molecule, epsilon associated protein; DNA-directed RNA polymerase I subunit RPA34; epsilon polypeptide associated protein; PAF49; RNA polymerase I-associated factor PAF49

Gene Aliases: ASE-1; ASE1; CAST; CD3EAP; PAF49

UniProt ID: (Human) O15446

Entrez Gene ID: (Human) 10849

Function(s)

transcription initiation from RNA polymerase I promoter transcription elongation from RNA polymerase I promoter termination of RNA polymerase I transcription transmembrane receptor protein tyrosine kinase signaling pathway rRNA transcription positive regulation of gene expression, epigenetic

Process(es)

DNA-directed RNA polymerase activity poly(A) RNA binding

PAF49 Polyclonal Antibody

Product Details

Product Specific Information

Target Information

Bioinformatics

Material safety data sheets (MSDS)

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