TREX1 Recombinant Rabbit Monoclonal Antibody [JG35-71]
Catalog# ET7108-16
TREX1 Recombinant Rabbit Monoclonal Antibody [JG35-71]
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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IP
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Human
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unconjugated
Overview
Product Name
TREX1 Recombinant Rabbit Monoclonal Antibody [JG35-71]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human TREX1 aa 110-314 / 314.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP
Target Molecular Weight
Predicted band size: 33 kDa
Positive Control
A431 cell lysate, SK-Br-3 cell lysate, Hela, LOVO, SiHa, human colon tissue, human prostate carcinoma tissue.
Conjugation
unconjugated
Clone Number
JG35-71
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000-1:2,000
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IF-Cell
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1:50-1:200
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IF-Tissue
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1:50-1:200
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IHC-P
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1:50-1:200
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IP
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1-2μg/sample
Target
Function
Major cellular 3'-to-5' DNA exonuclease which digests single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) with mismatched 3' termini. Prevents cell-intrinsic initiation of autoimmunity. Acts by metabolizing DNA fragments from endogenous retroelements, including L1, LTR and SINE elements. Unless degraded, these DNA fragments accumulate in the cytosol and activate the IFN-stimulatory DNA (ISD) response and innate immune signaling. Prevents chronic ATM-dependent checkpoint activation, by processing ssDNA polynucleotide species arising from the processing of aberrant DNA replication intermediates. Inefficiently degrades oxidized DNA, such as that generated upon antimicrobial reactive oxygen production or upon absorption of UV light. During GZMA-mediated cell death, contributes to DNA damage in concert with NME1. NME1 nicks one strand of DNA and TREX1 removes bases from the free 3' end to enhance DNA damage and prevent DNA end reannealing and rapid repair.
Background References
1. Gehrke N. et al. Oxidative damage of DNA confers resistance to cytosolic nuclease TREX1 degradation and potentiates STING-dependent immune sensing. Immunity 39:482-495(2013).
2. de Silva U. et al. The crystal structure of TREX1 explains the 3\' nucleotide specificity and reveals a polyproline II helix for protein partnering. J. Biol. Chem. 282:10537-10543(2007).
Sequence Similarity
Belongs to the exonuclease superfamily. TREX family.
Tissue Specificity
Detected in thymus, spleen, liver, brain, heart, small intestine and colon.
Post-translational Modification
Ubiquitinated, but not targeted to proteasomal degradation. Ubiquitination may be important for interaction with UBQLN1.
Subcellular Location
Cytoplasm, Endoplasmic reticulum, Membrane, Nucleus.
UNIPROT
Synonyms
3' 5' exonuclease TREX1 antibody
3' repair exonuclease 1 antibody
AGS1 antibody
AGS5 antibody
CRV antibody
Deoxyribonuclease III, dnaQ/mutD (E. coli) like antibody
DKFZp434J0310 antibody
DNase III antibody
DRN3 antibody
HERNS antibody
ExpandImages
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Western blot analysis of TREX1 on A431 cell lysates with Rabbit anti-TREX1 antibody (ET7108-16) at 1/1,000 dilution.
Lysates/proteins at 10 µg/Lane.
Predicted band size: 33 kDa
Observed band size: 33 kDa
Exposure time: 1 minute 50 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7108-16) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
ICC staining of TREX1 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7108-16, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of TREX1 in LOVO cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7108-16, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of TREX1 in SiHa cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7108-16, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-TREX1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-16, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue using anti-TREX1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-16, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Flow cytometric analysis of A431 cells labeling TREX1.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7108-16, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
TREX1 was immunoprecipitated from 0.2 mg A431 cell lysate with ET7108-16 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET7108-16 at 1/2,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: A431 cell lysate (input)
Lane 2: ET7108-16 IP in A431 cell lysate
Lane 3: Rabbit IgG instead of ET7108-16 in A431 cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"