Active Recombinant Human MAM Domain Containing Glycosylphosphatidylinositol Anchor 2, His-tagged

• Cat.No.: MDGA2-5687H
• Product Overview: Recombinant Human MDGA2protein was expressed in Murine myeloma cell line with a C-terminal 6-His tag. Gln21-Asp931.

Specification

• Species: Human
• Source: Mammalian Cells
• Tag: His
• Protein Length: 21-931 a.a.
• Description: MDGA2 is a 130 kDamember of the Ig superfamily of proteins. Human MDGA2 is synthesized as a 956amino acid (aa) precursor that contains a 25 aa signal sequence, a 906 aamature chain, and a 25 aa propeptide. The mature chain consists of sixIg-like domains, followed by a MAM domain (aa 746-921) and a GPI anchor. Inaddition, there are eight potential sites for N-linked glycosylation. Maturehuman MDGA2 shares 98% aa sequence identity with mature mouse and rat MDGA2.MDGA2 is structurally similar to other IgCAMS, such as the L1 family andaxonin 1, which have roles in cell adhesion, migration, and processoutgrowth.
• Form: Lyophilized from a0.2 μm filtered solution in PBS.
• N-terminalSequence: Gln21 predicted
• Molecular Weight: 103.2kDa
• Activity: Measuredby its ability to enhance neurite outgrowth of E18 rat embryonic corticalneurons. Able to significantly enhance neurite outgrowth when immobilized at7.5-30 μg on a nitrocellulose-coated microplate.
• Endotoxin Level: <1.0 EUper 1 μg of the protein by the LAL method.
• SDS-PAGE: 105-125kDa, reducing conditions.
• Purity: >90%,by SDS-PAGE under reducing conditions and visualized by silver stain.
• Reconstitution: 200μg/mL in sterile PBS
• Storage: Use amanual defrost freezer and avoid repeated freeze-thaw cycles. 12 months fromdate of receipt, -20 to -70°C as supplied. 1 month, 2 to 8°C under sterileconditions after reconstitution. 3 months, -20 to -70°C under sterileconditions after reconstitution.
• Publications:
  MDGA2 is a novel tumour suppressor cooperating with DMAP1 in gastric cancer and is associated with disease outcome. (2015)

Gene Information

• Gene Name: MDGA2 MAM domain containingglycosylphosphatidylinositol anchor 2 [ Homo sapiens ]
• Official Symbol: MDGA2
• Synonyms: MDGA2; MAM domaincontaining glycosylphosphatidylinositol anchor 2; MAMDC1; c14_5286; MAMdomain-containing glycosylphosphatidylinositol anchor protein 2; MAM domaincontaining 1; MAMDC1; OTTHUMP00000028200; OTTHUMP00000179128; MAMdomain-containing protein 1; UNQ8188/PRO23197
• Gene ID: 161357
• mRNA Refseq: NM_182830
• Protein Refseq: NP_878250
• MIM: 611128
• UniProt ID: Q7Z553
• Chromosome Location: 14q21.2

Citation

MDGA2 is a novel tumour suppressor cooperating with DMAP1 in gastric cancer and is associated with disease outcome

Journal: Gut    PubMed ID: 26206665    Data: 2016/10/1

Authors: Kunning Wang, Qiaoyi Liang, Jun Yu

Article Snippet:Purified recombinant human glutathione S-transferase (GST)-DMAP1 protein (Abnova, Taipei City, Taiwan) and His-tagged recombinant human MDGA2 (MDGA2-His; Creative BioMart, Shirley, New York, USA) were used for pull-down assay.. GST protein was expressed in E coli strain BL21 (DE3), and the GST protein was purified using a GSTrap column (GE Healthcare, Buckinghamshire, UK).GST protein was expressed in E coli strain BL21 (DE3), and the GST protein was purified using a GSTrap column (GE Healthcare, Buckinghamshire, UK).

MDGA2 is inactivated by promoter methylation in gastric cancer cell lines. (A) MDGA2 was silenced or downregulated in 10 out of 11 gastric cancer cell lines but readily expression in normal gastric tissue. The methylation status of MDGA2 was determined by methylation-specific PCR (MSP). M, methylated; U, unmethylated. (B) A typical CpG island is present at the promoter region of MDGA2. Each vertical bar represents a single CpG site. The transcription start site (TSS) is indicated by a curved arrow. A region for bisulfite genomic sequencing (BGS) and combined bisulfite restriction analysis (COBRA) is denoted. BGS analysis confirmed high levels of promoter methylation in MDGA2-silenced cells and no/low methylation in MDGA2-expressing samples. (C) MDGA2 mRNA expression was restored after treatment with the demethylation reagent 5-Aza. Decreased methylation was revealed by BGS after 5-Aza treatment.

Epigenetic inactivation of MDGA2 in primary gastric cancers. (A) Transcriptional downregulation of MDGA2 in gastric tumours compared with adjacent normal tissues was shown by reverse transcription PCR (RT-PCR). (B) Protein levels of MDGA2 were quantitated as the percentage of cells with positive staining by immunohistochemistry in 20 pairs of gastric tumour and adjacent non-tumour samples. (C) Methylation-specific PCR (MSP) showed high methylation of MDGA2 promoter in 10 gastric tumours and low methylation in their adjacent non-tumour tissues, which was confirmed by bisulfite genomic sequencing (BGS). M, methylated; U, unmethylated. (D) Kaplan–Meier curves showed that MDGA2 methylation is significantly associated with shortened survival in overall patients with gastric cancer and (E) in patients at an early stage but not patients at late stages (log rank test).

Univariate and multivariate Cox regression analyses of potential poor prognostic factors in gastric cancer