Invitrogen

PAX3 Recombinant Polyclonal Antibody (16HCLC)

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Datasheet

Protocols

Questions & Answers

Tech Support

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PAX3 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of PAX3 in HeLa cells using a PAX3 Recombinant Rabbit Polyclonal Antibody (Product # 710178) followed by detection using an Alexa Fluor 488-conjugated Goat anti-Rabbit secondary antibody (green) (Image A). Nuclei were stained using DAPI (Image B) and actin stained with Alexa Fluor 594 phalloidin (red) (image C). Image D is a composite image showing nuclear localization of Pax 3.

PAX3 Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts of HeLa (Lane 1) and A431 (Lane 2). The blot was probed with Anti-PAX3 Recombinant Rabbit Polyclonal Antibody (Product # 710178, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 53 kDa band corresponding to PAX3 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

PAX3 Antibody in ChIP assay (ChIP) — Chromatin immunoprecipitation analysis of Pax3 was performed using cross-linked chromatin from 1 x 10^6 HCT116 human colon carcinoma cells treated with serum for 0, 15, and 60 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 Pax3 Recombinant Rabbit Polyclonal Antibody (Product # 710178). Chromatin aliquots from ~1 x 10^5 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-3.2kb upstream of human FOS, exon-4 of FOS, or the imprinting control region (ICR) of the human H19 locus. 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 representation of human FOS and H19 loci are shown above the data where boxes represent exons (grey boxes = translated regions, white boxes = untranslated regions), the zigzag lines represent introns, and the straight lines represent upstream sequences. Regions amplified by FOS and H19 primers are represented by black bars. Data courtesy of the Innovators Program.

Product Details

710178

Applications	Tested Dilution	Publications
Western Blot (WB)	2-4 µg/mL	-
Immunocytochemistry (ICC/IF)	1-3 µg/mL	-
ChIP assay (ChIP)	1 µL	-

Product Specifications
Species Reactivity	Human
Host/Isotype	Rabbit / IgG
Class	Recombinant Polyclonal
Type	Antibody
Clone	16HCLC
Immunogen	Recombinant protein corresponding to amino acids 34–276 of human paired box 3 View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Protein A
Storage buffer	PBS
Contains	0.09% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2532605

Product Specific Information

This antibody is predicted to react with mouse based on sequence homology.

Recombinant rabbit polyclonal antibodies are unique offerings from Thermo Fisher Scientific. They are comprised of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds - the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies - all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody - recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets - a recombinant rabbit polyclonal antibody has a known mixture of light and heavy chains. The exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.

Target Information

Pax 3 (Paired Box 3) is a transcription factor that is involved in development of the peripheral nervous system, melanocytes, some vascular smooth muscle, and a number of other derivatives. It regulates neurogenesis in pre-migratory neural crest cells from the dorsal neural tube, and in myogenic progenitors in the presomitic mesoderm and the hypaxial somites. Alternative splicing results in isoforms having different C-termini.

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

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Bioinformatics

Protein Aliases: HuP2; MGC120381; MGC120382; MGC120383; MGC120384; MGC134778; paired box homeotic gene 3; Paired box protein Pax-3; paired domain gene 3; paired domain gene HuP2

Gene Aliases: CDHS; HUP2; PAX3; WS1; WS3

UniProt ID: (Human) P23760

Entrez Gene ID: (Human) 5077

Function(s)

transcription from RNA polymerase II promoter apoptotic process nervous system development muscle organ development sensory perception of sound organ morphogenesis neural crest formation positive regulation of transcription, DNA-templated positive regulation of transcription from RNA polymerase II promoter

Process(es)

transcription factor activity, sequence-specific DNA binding protein binding sequence-specific DNA binding HMG box domain binding