Recombinant Rat SPI1 Protein

• Cat.No.: SPI1-5706R
• Product Overview: Recombinant Rat SPI1 full length or partial length protein was expressed.

Specification

• Species: Rat
• Source: Mammalian Cells
• Tag: His
• Form: Liquid or lyophilized powder
• Endotoxin: < 1.0 EU per μg of the protein as determined by the LAL method.
• Purity: >80%
• Notes: This item requires custom production and lead time is between 5-9 weeks. We can custom produce according to your specifications.
• Storage: Store it at +4 ºC for short term. For long term storage, store it at -20 ºC~-80 ºC.
• Storage Buffer: PBS buffer

Gene Information

• Gene Name: Spi1 spleen focus forming virus (SFFV) proviral integration oncogene spi1 [ Rattus norvegicus ]
• Official Symbol: SPI1
• Gene ID: 366126
• mRNA Refseq: NM_001005892.2
• Protein Refseq: NP_001005892.1
• UniProt ID: Q6BDS1

Citation

Open Chromatin Profiling in Adipose Tissue Marks Genomic Regions with Functional Roles in Cardiometabolic Traits

Journal: G3: Genes|Genomes|Genetics    PubMed ID: 31186305    Data: 2019/6/11

Authors: Maren E. Cannon, Kevin W. Currin, Karen L. Mohlke

Article Snippet:For EMSA, we prepared nuclear cell extracts from SGBS preadipocyte and SW872 cells using the NE-PER nuclear and cytoplasmic extraction kit (Thermo Scientific) as previously described. ( Kulzer et al. 2014 ) Double-stranded oligos (Table S12) were incubated with SGBS preadipocyte or SW872 nuclear extract or 100 ng purified PU.1 protein (Creative BioMart SPI1-172H) and DNA-protein complex visualization was carried out as previously described. ( Kulzer et al. 2014 ) A positive control oligo contained the PU.1 motif from JASPAR and a negative control did not contain the motif (Table S12).. We repeated all EMSA experiments on independent days and obtained consistent results.We repeated all EMSA experiments on independent days and obtained consistent results.

A variant at the ATP2A1-SH2B1 BMI GWAS locus alters chromatin accessibility and PU.1 binding. ( A) rs7187776 is located in the promoter of a long SH2B1 isoform, transcribed left-to-right in the image; the 5′-UTR of TUFM , transcribed right-to-left in the image; and a region containing ATAC-seq peaks from multiple sources. ETS1 and PU.1 ENCODE ChIP-seq binding was observed in K562 and GM12891, respectively. Many additional transcription factor ChIP-seq peaks overlap this region in the ENCODE datasets. (B) A 19-nt probe containing rs7187776-G shows increased protein binding to purified PU.1 in EMSA, similar to a positive control probe containing the consensus PU.1 motif (+). A negative control probe (-) and a probe containing rs7187776-A showed no binding to PU.1. Black arrows indicate allele-specific protein binding, gray arrow indicates the well of the gel. Similar protein binding patterns and equal amounts of free DNA probe were observed using SW872 nuclear extract (Figure S5). PU.1 consensus motif from JASPAR ( Mathelier et al. 2016 ). (C) The genomic region containing rs7187776-A shows increased transcriptional activity and allelic differences in THP-1 monocytes. The genomic region including part of the 3′ UTR of TUFM and part of the 5′ UTR of SH2B1 was cloned upstream of a minimal promoter and the luciferase gene. Dots represent the average of two technical replicates. Forward and reverse designated with respect to the genome, so forward corresponds to left-to-right in the image. P-values determined by Student’s t -test. EV, empty vector. (D) Summary of the direction of effect of rs7187776-G.