Recombinant Human E3 Ubiquitin-Protein Ligase Siah1 (SIAH1) Protein (GST)

Name: Recombinant Human E3 Ubiquitin-Protein Ligase Siah1 (SIAH1) Protein (GST)
Brand: Beta LifeScience
SKU: BLC-03744P
Availability: InStock

Greater than 90% as determined by SDS-PAGE.

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Product Overview

Description	Recombinant Human E3 Ubiquitin-Protein Ligase Siah1 (SIAH1) Protein (GST) is produced by our E.coli expression system. This is a full length protein.
Purity	Greater than 90% as determined by SDS-PAGE.
Uniprotkb	Q8IUQ4
Target Symbol	SIAH1
Synonyms	E3 ubiquitin-protein ligase SIAH1; FLJ08065; hSIAH1; HUMSIAH; Seven in absentia homolog 1 (Drosophila); Seven in absentia homolog 1; Siah 1; Siah 1a; Siah E3 ubiquitin protein ligase 1; Siah-1; Siah-1a; Siah1; SIAH1_HUMAN; SIAH1A; Ubiquitin ligase SIAH1
Species	Homo sapiens (Human)
Expression System	E.coli
Tag	N-GST
Target Protein Sequence	MTGKATPPSLYSWRGVLFTCLPAARTRKRKEMSRQTATALPTGTSKCPPSQRVPALTGTTASNNDLASLFECPVCFDYVLPPILQCQSGHLVCSNCRPKLTCCPTCRGPLGSIRNLAMEKVANSVLFPCKYASSGCEITLPHTEKADHEELCEFRPYSCPCPGASCKWQGSLDAVMPHLMHQHKSITTLQGEDIVFLATDINLPGAVDWVMMQSCFGFHFMLVLEKQEKYDGHQQFFAIVQLIGTRKQAENFAYRLELNGHRRRLTWEATPRSIHEGIATAIMNSDCLVFDTSIAQLFAENGNLGINVTISMC
Expression Range	1-313aa
Protein Length	Full Length of Isoform 2
Mol. Weight	61.6kDa
Research Area	Neuroscience
Form	Liquid or Lyophilized powder
Buffer	Liquid form: default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol. Lyophilized powder form: the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose, pH 8.0.
Reconstitution	Briefly centrifuged the vial prior to opening to bring the contents to the bottom. Reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL. It is recommended to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. The default final concentration of glycerol is 50%.
Storage	1. Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. 2. Avoid repeated freeze-thaw cycles. 3. Store working aliquots at 4°C for up to one week. 4. In general, protein in liquid form is stable for up to 6 months at -20°C/-80°C. Protein in lyophilized powder form is stable for up to 12 months at -20°C/-80°C.
Notes	Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Target Details

Target Function	E3 ubiquitin-protein ligase that mediates ubiquitination and subsequent proteasomal degradation of target proteins. E3 ubiquitin ligases accept ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then directly transfers the ubiquitin to targeted substrates. Mediates E3 ubiquitin ligase activity either through direct binding to substrates or by functioning as the essential RING domain subunit of larger E3 complexes. Triggers the ubiquitin-mediated degradation of many substrates, including proteins involved in transcription regulation (ELL2, MYB, POU2AF1, PML and RBBP8), a cell surface receptor (DCC), the cell-surface receptor-type tyrosine kinase FLT3, the cytoplasmic signal transduction molecules (KLF10/TIEG1 and NUMB), an antiapoptotic protein (BAG1), a microtubule motor protein (KIF22), a protein involved in synaptic vesicle function in neurons (SYP), a structural protein (CTNNB1) and SNCAIP. Confers constitutive instability to HIPK2 through proteasomal degradation. It is thereby involved in many cellular processes such as apoptosis, tumor suppression, cell cycle, axon guidance, transcription regulation, spermatogenesis and TNF-alpha signaling. Has some overlapping function with SIAH2. Induces apoptosis in cooperation with PEG3. Upon nitric oxid (NO) generation that follows apoptotic stimulation, interacts with S-nitrosylated GAPDH, mediating the translocation of GAPDH to the nucleus. GAPDH acts as a stabilizer of SIAH1, facilitating the degradation of nuclear proteins. Mediates ubiquitination and degradation of EGLN2 and EGLN3 in response to the unfolded protein response (UPR), leading to their degradation and subsequent stabilization of ATF4.
Subcellular Location	Cytoplasm. Nucleus. Note=Predominantly cytoplasmic. Partially nuclear.
Protein Families	SINA (Seven in absentia) family
Database References	HGNC: 10857 OMIM: 602212 KEGG: hsa:6477 STRING: 9606.ENSP00000349156 UniGene: PMID: 28843785 The crystallographic models provide structural insights into the substrate binding of the SIAH family E3 ubiquitin ligases that are critically involved in regulating cancer-related pathways. PMID: 27776223 Results showed that SIAH1 and PTP4A1 expression was regulated by mir-944 in breast cancer cells. miR-944 binds directly the 3 UTR of their promotor region. PMID: 27377268 Study identified SIAH1/2 (SIAH) as the E3 ligase mediating Wnt-induced Axin degradation. SIAH proteins promote the ubiquitination and proteasomal degradation of Axin through interacting with a VxP motif in the GSK3-binding domain of Axin, and this function of SIAH is counteracted by GSK3 binding to Axin. PMID: 28546513 results suggest that CacyBP/SIP is involved in regulation of the Hsp90 chaperone machinery PMID: 27249023 PINK1 disease mutants failed to recruit synphilin-1 and did not activate mitophagy, indicating that PINK1-synphilin-1-SIAH-1 represents a new parkin-independent mitophagy pathway. Drugs that activate this pathway will provide a novel strategy to promote the clearance of damaged mitochondria in Parkinson's disease. PMID: 27334109 Results suggest that activated STAT3 regulates active beta-catenin protein levels via stabilization of SIAH-1 and the subsequent ubiquitin-dependent proteasomal degradation of beta-catenin in HEK293T cells. PMID: 27871173 Siah-1 expression was observed in 41.4% of oral squamous cell carcinoma. Siah-1 was expressed in the cytoplasm and cell membranes, and partially in the nucleus . PMID: 27616748 SIAH1 is associated with a tumor promoting role in breast cancer. PMID: 26654769 In neuroblastoma cells, siRNA silencing of SIAH1 promoted cellular proliferation and suppressed apoptosis. Protein and mRNA expression of alpha-synuclein, LC3-II and p53 decreased after SIAH1 knockdown. E1 protein and mRNA levels increased after SIAH1 siRNA. PMID: 26722480 Results demonstrated that miR-107 directly down-regulated SIAH1 expression in human breast cancer cells. An inverse correlation between the expression of miR-107 and SIAH1 was found in human breast cancer tissues and cell lines. PMID: 25851994 Similar to TAp73, DNp73 is stabilized by hypoxia in a HIF-1a-dependent manner, which otherwise is degraded by Siah1. PMID: 26267146 findings demonstrate that dissociation of the GAPDH/Siah1 pro-apoptotic complex can block high glucose-induced pericyte apoptosis, widely considered a hallmark feature of DR PMID: 26438826 Data indicate that ubiquitin ligase Siah as a regulator of to Mesenchymal Transition (EMT) by controlling the abundance of the key transcription factor Zeb1, while Siah itself is subject to regulation by EMT-inducing factors. PMID: 25528765 miRNA network regulates SIAH1 in in colorectal metastasis tissues. PMID: 26069251 CDK2 up-regulates the protein level of KLF10 through reducing its association with SIAH1, a KLF10 E3-ubiqutin ligase involved in proteasomal degradation. PMID: 25728284 molecular basis of Siah1 and Siah2 E3 ubiquitin ligase substrate specificity PMID: 25202994 Siah1 is a substrate of ASK1 for activation of the GAPDH-Siah1 oxidative stress signaling cascade. PMID: 25391652 SIAH-1 plays an essential role in beta-catenin degradation in HCT116 CRC cells and that CSN5 affects beta-catenin target gene expression in these cells. PMID: 24882689 miR-135a/SIAH1/beta-catenin signaling is important in the transformation and progression of cervical carcinoma. PMID: 24503442 The spatial orientation of one zinc-finger subdomain in Siah1 ubiquitin ligase differs with respect to the rest of the dimeric protein. PMID: 24316825 the molecular mechanisms modulating ACK1 PMID: 23208506 USP19 interacts with the ubiquitin ligases SIAH1 and SIAH2, which promote USP19 ubiquitylation and degradation by the proteasome. PMID: 23500468 High expression of seven-in-absentia homologue 1 is associated with nasopharyngeal carcinoma. PMID: 23228635 Calmodulin-regulated Siah-1A binding to mGluR5 dynamically regulates mGluR5 trafficking PMID: 23152621 Siah1 is the E3 ubiquitin ligase for ELL2 polyubiquitination and proteasomal degradation. PMID: 22483617 In sharp contrast to SIAH1/SIAH2 and UBCH8, TRIAD1 binding to PML-RARalpha has no effect on its turnover. PMID: 22037423 Taken together, our data suggest EFF1D functions as a novel negative regulator of SIAH-1. PMID: 21633900 Siah-1 may play an important role in ubiquitin-dependent degradation of hepatitis B virus X protein and may be involved in suppressing the progression of hepatocellular carcinoma (HCC). PMID: 21878328 infection of SIAH-1 knockdown cells with HSV, higher levels of ICP0 were produced, ICP0 was less ubiquitinated, and the half-life of this multifunctional viral regulatory protein was increased PMID: 21632771 results suggest that hypoxia downregulates beta-catenin by increasing Siah-1 expression in a p53-dependent manner PMID: 21466614 ARTS interacts with the E3 ligase Siah-1 (seven in absentia homolog 1) to induce ubiquitination and degradation of XIAP. PMID: 21185211 Herp temporarily bound to E3 ligase SIAH1a during proteolytic stress but not during ER stress. PMID: 20604806 Overexpression of Siah1L and Siah1 is associated with radiosensitivity of breast cancer. PMID: 20682032 these results propose a novel role of SIAH-1 in regulating the expression level of HPH2 through the ubiquitin-proteasome pathway. PMID: 20471960 Siah1 is a bona fide E2F1 target gene, which at least partly, mediates the suppression of beta-catenin/TCF signalling pathway PMID: 20187294 Immunofluorescence microscopy shows that the intracellular distribution of SIAH-1 and Kid/KIF22 appears to be modified in human tumor tissues compared to normal controls PMID: 20144232 increase of the function of SIAH1 to upregulate the expression of Bim may play an important role in the progression of breast cancer PMID: 20082325 Direct ubiquitination of beta-catenin by Siah-1 and regulation by the exchange factor TBL1. PMID: 20181957 JNK and ERK signaling pathways may play an important role in the SIAH1-dependent biological behavior of breast cancer. PMID: 19775288 siah-1 Protein is necessary for high glucose-induced glyceraldehyde-3-phosphate dehydrogenase nuclear accumulation and cell death in Muller PMID: 19940145 Siah proteins function as E3 ubiquitin-protein ligases PMID: 11786535 The Siah-1 gene promoter was cloned and the basic structure of the Siah-1 gene was determined. PMID: 11852084 genetic regulation in tumor reversion PMID: 12399545 structural analysis and interaction with Siah-interacting protein PMID: 12421809 SIAH1 has been identified as a putative tumor suppressor gene for human hepatocellular carcinomas, with a correlation between its suppressed expression and tumor size and differentiation. PMID: 12557228 PEG10 protein associated with SIAH1, a mediator of apoptosis, and overexpression of PEG10 decreased the cell death mediated by SIAH1. PMID: 12810624 Siah-1 was found to abrogate the inhibitory effects of synphilin-1 on dopamine release PMID: 14506261 A p53-binding site was identified in the siah-1b promoter, located at nucleotides -2155/-2103 relative to the translational start site. PMID: 14985507 SIAH-mediated down regulation of alternative splicing may be an important developmental difference between otherwise highly conserved T-STAR proteins. PMID: 15163637

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Proteins are sensitive to heat, and freeze-drying can preserve the activity of the majority of proteins. It improves protein stability, extends storage time, and reduces shipping costs. However, freeze-drying can also lead to the loss of the active portion of the protein and cause aggregation and denaturation issues. Nonetheless, these adverse effects can be minimized by incorporating protective agents such as stabilizers, additives, and excipients, and by carefully controlling various lyophilization conditions.

Commonly used protectant include saccharides, polyols, polymers, surfactants, some proteins and amino acids etc. We usually add 8% (mass ratio by volume) of trehalose and mannitol as lyoprotectant. Trehalose can significantly prevent the alter of the protein secondary structure, the extension and aggregation of proteins during freeze-drying process; mannitol is also a universal applied protectant and fillers, which can reduce the aggregation of certain proteins after lyophilization.

Our protein products do not contain carrier protein or other additives (such as bovine serum albumin (BSA), human serum albumin (HSA) and sucrose, etc., and when lyophilized with the solution with the lowest salt content, they often cannot form A white grid structure, but a small amount of protein is deposited in the tube during the freeze-drying process, forming a thin or invisible transparent protein layer.

Reminder: Before opening the tube cap, we recommend that you quickly centrifuge for 20-30 seconds in a small centrifuge, so that the protein attached to the tube cap or the tube wall can be aggregated at the bottom of the tube. Our quality control procedures ensure that each tube contains the correct amount of protein, and although sometimes you can't see the protein powder, the amount of protein in the tube is still very precise.

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