货号:03-120
品牌:Merck Millipore
规格:
目录价:¥4926.00
市场价格:¥4187.10
会员价格:¥3940.80
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Description:
RIPAb+™ Aly/REF
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Promotional Text:
Special Shipping Offer on Antibodies100% Performance Guaranteed
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Product Overview:
RIPAb+ antibodies are evaluated using the RNA Binding Protein Immunoprecipitation (RIP) assay. Each RIPAb+ antibody set includes a negative control antibody to ensure specificity of the RIP reaction and is verified for the co-immunoprecipitation of RNA associated specifically with the immunoprecipitated RNA binding protein of interest. Where appropriate, the RIPAb+ set also includes quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully co-precipitating the specific RNA targets, such as messenger RNAs. The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. If a target specific assay is not provided, the RIPAb+ kit is validated using an automated microfluidics-based assay by enrichment of detectable RNA over control immunoprecipitation.Aly/REF (THO complex subunit 4, THOC4) is a heat stable, nuclear protein that functions as a molecular chaperone. Aly/REF is preferentially associated with intronless mRNAs. Specific protein complexes, exon junction complexes and transcription/export complexes, are loaded onto mRNAs in a splicing-dependent manner, and adaptor proteins, such as Aly, in turn recruit mRNA exporters onto the RNA.
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Specificity:
This antibody recognizes Aly/REF.
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Molecular Weight:
~ 32 kDa
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Immunogen:
Recombinant protein corresponding to human Aly/REF.
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Isotype:
IgG1κ
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Background Information:
Aly (THO complex subunit 4 or THOC4 or Aly/REF) is a heat stable, nuclear protein that functions as a molecular chaperone. It is thought to regulate dimerization, DNA binding, and transcriptional activity of basic region-leucine zipper (bZIP) proteins. Aly (THOC4) is also preferentially associated with intronless mRNAs in the nucleus. Loading of export factors onto mRNAs is a key step in gene expression. In vertebrates, splicing plays a role in this process. Specific protein complexes, exon junction complexes and transcription/export complexes, are loaded onto mRNAs in a splicing-dependent manner, and adaptor proteins such as Aly (THOC4) in the complexes in turn recruit mRNA exporters onto the RNA.
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Species Reactivity:
Human
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Species Reactivity Note:
Demonstrated to react with human.
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Application Notes:
Immunoprecipitation from RIP lysate:Representative lot data.RIP lysate from HeLa cells (~2 X 10E7 cell equivalents per IP) was subjected to immunoprecipitation using either 5 µg of a normal mouse IgG, (Cat. #CS200621), or 5 µg of Anti-Aly/REF antibody (Cat. # CS204402). Ten percent of the precipitated proteins (lane 1: mouse IgG, lane 3: Aly/REF) and HeLa whole cell lysate (lane 2) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Aly/REF antibody (Cat. # CS204402, 1:1000). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00232).Arrow indicates Aly/REF (~32 kDa) (Figure 2).Western Blot Analysis: Representative lot data.Hek293 cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with (0.5ug/mL) dilution.Proteins were visualized using a Goat Anti-Mouse IgG conjugated to HRP using a chemiluminescence detection system.Arrow indicates Aly/REF (~32 kDa) (Figure 3).Immunocytochemistry Analysis: A 1:500 dilution from a representative lot detected Aly/REF (red) in NIH/3T3 and HeLa cells. The antibody positively stains the nucleus (Figure 4).Immunoprecipitation Analysis: A representative lot was used by an independent laboratory in IP. (Kim, V.N., et al. (2001). The EMBO Journal. 20, 2062-2068.)
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Control:
Includes negative control mouse IgG antibody and control primers specific for the mRNA of human DHFR-1.
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Quality Assurance:
RNA Binding Protein Immunoprecipitation:RIP Lysate prepared from Jurkat cells (~5 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using either 5 µg of a normal mouse IgG (Cat. # CS200621), or 5 µg of Anti-Aly/REF antibody (Cat. # CS204402) and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).Successful immunoprecipitation of Aly/REF-associated RNA was verified by qPCR using RIP Primers DHFR-1, (Cat. # CS204401) (Figure 1). Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.
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Purification Method:
Protein G purfied
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Presentation:
Anti-Aly/REF (Mouse Monoclonal), Part # CS204402. One vial containing 50 µg of protein G purified monoclonal antibody in buffer containing 0.1 M Tris-glycine, 0.105 M NaCl, pH 7.4, 0.05% sodium azide, before the addition of 30% glycerol. Store at -20°C.Normal Mouse IgG, Part # CS200621. One vial containing 125 µg purified mouse IgG in 125 µL storage buffer containing 0.1% sodium azide. Store at -20°C.RIP Primers, DHFR-1, Part # CS204401. One vial containing 75 μL of 5 μM of each primer specific for human DHFR-1 mRNA. Store at -20°C.FOR: TTC CTG AGA AGA ATC GAC CTT TAA AREV: AAG GCA TCA TCT AGA CTT CTG GAA A
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Storage Conditions:
Stable for 1 year at -20°C from date of receipt.Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
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UniProt Number:
Q86V81
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Entrez Gene Number:
NP_005773.3
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Gene Symbol:
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ALY
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BEF
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Tho4
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THOC4
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Alternate Names:
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Ally of AML-1 and LEF-1
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THO complex 4
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Transcriptional coactivator Aly/REF
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bZIP enhancing factor
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bZIP-enhancing factor BEF
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transcriptional coactivator
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Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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Key Applications:
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Western Blotting
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Immunocytochemistry
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RNA Binding Protein Immunoprecipitation (RIP)
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Entrez Gene Summary:
The protein encoded by this gene is a heat stable, nuclear protein and functions as a molecular chaperone. It is thought to regulate dimerization, DNA binding, and transcriptional activity of basic region-leucine zipper (bZIP) proteins. [provided by RefSeq].
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UniProt Summary:
FUNCTION: Acts as chaperone and promotes the dimerization of transcription factors containing basic leucine zipper (bZIP) domains and thereby promotes transcriptional activation. Plays a role in mRNA processing and export. May function as scaffold that mediates interactions between proteins and/or RNA. Integral part of the THO/TREX complex that is recruited to transcribed genes and travels with the RNA polymerase during elongation. Is part of the exon junction complex that remains associated with spliced mRNA and plays an important role in mRNA export and nonsense-mediated RNA decay. Directs mRNA derived from Herpes simplex virus intron-less genes to the NXF1-mediated export pathway.SUBUNIT STRUCTURE: Homomultimer. Binds LEF1 and RUNX1 By similarity. Is part of several complexes involved in mRNA processing and export. Part of the heteromultimeric THO/TREX complex containing THOC1, THOC2, THOC3, THOC4 and UAP56. Associates with the spliceosome. Binds NXF1 and RBM8A directly and is part of the exon junction complex (EJC) containing NCBP1, NCBP2, RBM8A, SRRM1, NXF1, RENT2, RENT3B and THOC4 By similarity. Found in a mRNA splicing-dependent exon junction complex (EJC) with DEK, RBM8A, RNPS1, SRRM1 and THOC4. Binds UL54 from Herpes simplex type 1 (strain KOS). Found in a mRNP complex with RENT3A and RENT3B. Interacts with BAT1, RBM8A, RNPS1 and SRRM1. Identified in the spliceosome C complex, at least composed of AQR, ASCC3L1, C19orf29, CDC40, CDC5L, CRNKL1, DDX23, DDX41, DDX48, DDX5, DGCR14, DHX35, DHX38, DHX8, EFTUD2, FRG1, GPATC1, HNRPA1, HNRPA2B1, HNRPA3, HNRPC, HNRPF, HNRPH1, HNRPK, HNRPM, HNRPR, HNRPU, KIAA1160, KIAA1604, LSM2, LSM3, MAGOH, MORG1, PABPC1, PLRG1, PNN, PPIE, PPIL1, PPIL3, PPWD1, PRPF19, PRPF4B, PRPF6, PRPF8, RALY, RBM22, RBM8A, RBMX, SART1, SF3A1, SF3A2, SF3A3, SF3B1, SF3B2, SF3B3, SFRS1, SKIV2L2, SNRPA1, SNRPB, SNRPB2, SNRPD1, SNRPD2, SNRPD3, SNRPE, SNRPF, SNRPG, SNW1, SRRM1, SRRM2, SYF2, SYNCRIP, TFIP11, THOC4, U2AF1, WDR57, XAB2 and ZCCHC8.SUBCELLULAR LOCATION: Nucleus. Cytoplasm. Note: Travels to the cytoplasm as part of the exon junction complex (EJC) bound to mRNA. PTM: Arg-50 and Arg-204 are dimethylated, probably to asymmetric dimethylarginine. Phosphorylated upon DNA damage, probably by ATM or ATR. INVOLVEMENT IN DISEASE: Antibodies against THOC4 are found in sera of patients with systemic lupus erythematosus (SLE). SEQUENCE SIMILARITIES: Contains 1 RRM (RNA recognition motif) domain.
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Product Name:
RIPAb+™ Aly/REF
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Concentration:
0.7 mg/mL
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Antibody Type:
Monoclonal Antibody
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Qty/Pk:
10 assays
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Format:
Purified
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Host:
Mouse
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Packaging:
10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).
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