Recombinant Human PAX7

• Cat.No.: PAX7-29057TH
• Product Overview: Recombinant fragment of Human PAX7 with N terminal proprietary tag, 38.21kDa inclusive of tag.

Specification

• Species: Human
• Source: Wheat Germ
• Tag: Non
• Protein Length: 111 amino acids
• Description: This gene is a member of the paired box (PAX) family of transcription factors. Members of this gene family typically contain a paired box domain, an octapeptide, and a paired-type homeodomain. These genes play critical roles during fetal development and cancer growth. The specific function of the paired box 7 gene is unknown but speculated to involve tumor suppression since fusion of this gene with a forkhead domain family member has been associated with alveolar rhabdomyosarcoma. Three transcript variants encoding different isoforms have been found for this gene.
• Molecular Weight: 38.210kDa inclusive of tags
• Form: Liquid
• Purity: Proprietary Purification
• Storage buffer: pH: 8.00Constituents:0.3% Glutathione, 0.79% Tris HCl
• Storage: Shipped on dry ice. Upon delivery aliquot and store at -80oC. Avoid freeze / thaw cycles.
• Sequences of amino acids: GGLDSATSISASCSQRADSIKPGDSLPTSQAYCPPTYSTT GYSVDPVAGYQYGQYGQSECLVPWASPVPIPSPTPRASCL FMESYKVVSGWGMSISQMEKLKSSQMEQFT
• Sequence Similarities: Belongs to the paired homeobox family.Contains 1 homeobox DNA-binding domain.Contains 1 paired domain.

Gene Information

• Gene Name: PAX7 paired box 7 [ Homo sapiens ]
• Official Symbol: PAX7
• Synonyms: PAX7; paired box 7; paired box gene 7; paired box protein Pax-7; Hup1
• Gene ID: 5081
• mRNA Refseq: NM_001135254
• Protein Refseq: NP_001128726
• MIM: 167410
• Uniprot ID: P23759
• Chromosome Location: 1p36.13
• Pathway: Transcriptional misregulation in cancers, organism-specific biosystem; Transcriptional misregulation in cancers, conserved biosystem
• Function: sequence-specific DNA binding; sequence-specific DNA binding transcription factor activity

Citation

DNA aptamers against the DUX4 protein reveal novel therapeutic implications for FSHD

Journal: The FASEB Journal    PubMed ID: 32020675    Data: 2020/2/5

Authors: Christian Klingler, Jon Ashley, Jochen Kinter

Article Snippet:Aptamers were compared against 5′‐Atto647N labeled aptamers as probes with sequences described in in Supplemental Table T1.Aptamers were compared against 5′‐Atto647N labeled aptamers as probes with sequences described in in Supplemental Table T1.. The labeled aptamers were incubated with 1 nM of either StrepII‐SUMO‐DUX4‐His6, 1 nM of PAX7‐His6 (Creative BioMart, Shirley US‐NA), or 15 nM of PROP1‐cMyc‐DDK(FLAG) (OriGene Technologies, Inc, Rockville, US‐MD) recombinant proteins and various concentrations of test aptamers (Microsynth, Balgach, Switzerland) in an assay buffer containing 50 mM of Tris‐HCl pH 7.5, 150 mM of NaCl, 5 mM of MgCl 2 , 0.1% of BSA, 0.1% of Triton X‐100, 1 mM of DTT, and 0.35 nM of MAb anti‐6His Terbium cryptate Gold antibody (cisbio, Codolet, France) in white 384 well microplates (Greiner Bio‐one, Kremsmünster, Austria) for 24 hours at 4°C.. Donor fluorescence (340 nm/620 nm) and acceptor fluorescence (340 nm/665 nm) was quantified on a TECAN Spark microplate reader with a lag time of 100 μs and integration time of 300 μs at 37°C.Donor fluorescence (340 nm/620 nm) and acceptor fluorescence (340 nm/665 nm) was quantified on a TECAN Spark microplate reader with a lag..

Testing the binding of SELEX‐originated aptamers. Comparing top nine list of SELEX generated aptamers in TR‐FRET assays. Aptamers were compared by competing against the 5′‐end Atto647N‐labeled template aptamer after incubation with StrepII‐SUMO‐DUX4‐His6. The fitted curves are displayed together with the IC50 values in micromolar from single experiments with ±95% confidence interval (bar plot). The SELEX aptamers were sorted according to their cluster size in counts‐per‐million (cpm) based on AptaSUITE software preprocessing steps (values in brackets). NA are not‐fittable data points indicating IC50 values higher than 10 μM

Specificity of DUX4 aptamers. A, Different aptamers were tested against the consensus PROP1 binding sequence inserted into the aptamer backbone by means of a TR‐FRET competition assays. Single experiment data is shown as IC50 values of various aptamers ± standard error of the data fitting to a four‐parameter logistic using R package “drc” (curves are provided in Supplemental Figures A‐C). B, TR‐FRET competition assay was performed to compare the binding of different aptamers to PAX7. C, TR‐FRET competition assay was performed to compare the binding of different aptamers to DUX4. D, DNA target sequence logos of PAX7 (JASPAR ID: MA0680.1) and PROP1 (JASPAR ID: MA0715.1) extracted from JASPAR database are displayed. E, DNA sequences of all aptamers used for this experiment are listed. Target motifs are depicted in bold font. Special sequence alterations are given in red font and bulge loops in blue font. F, The affinity gain caused by the affinity bulge loops in the DUX4 aptamer were compared in the presence of different transcription factors. The IC50 ratios (ordinate) between aptamers DUX4_CTT and DUX4_CTT_loop from TR‐FRET competition assays were compared. Different concentrations of aptamers were incubated with the recombinant proteins DUX4, PAX7, and PROP1 (abscissa) and corresponding Atto647N‐labeled probes