ASK1 Antibody (YA609)
(Synonyms: ASK1, MAPKKK5, MEKK5, MAP3K5, Mitogen-activated protein kinase kinase kinase 5, Apoptosis signal-regulating kinase 1, MAPK/ERK kinase kinase 5, ASK-1, MEK kinase 5, MEKK 5)Based on 1 Customer Validation
ASK1 Antibody (YA609) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to ASK1.
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Host:
Rabbit
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Isotype:
IgG
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Application:
WB, ICC/IF, IHC-P, FC
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Reactivity :
Human, Mouse
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Formulation:
Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.
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Conjugation:
Non-conjugated
Applications
| Application |
WB
WB: Western Blot
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ICC/IF
ICC/IF: Immunocytochemistry/
Immunofluorescence |
IHC-P
IHC-P: Immunohistochemistry-Paraffin
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FC
FC: Flow Cytometry
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| Dilution Ratio | 1:500-1:2000 | 1:50-1:200 | 1:50-1:200 | 1:50-1:100 |
Product Details
ASK1 Antibody (YA609) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to ASK1.
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Host Rabbit
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Clonality Recombinant,Monoclonal
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Species ReactivityHuman, Mouse
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Observed Molecular WeightObserved band size: 150 kDaNote: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
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Calculated Molecular Weight Predicted band size: 155 kDa
Entrez Gene: 4217 Human ; 26408 Mouse ; 365057 Rat
SwissProt: Q99683 Human ; O35099 Mouse ;
OMIM: 602448 Human
Synthetic peptide within N-terminal human ASK1.
Endogenous
Protein A affinity purified.
Non-conjugated
Unmodified
IgG
Product Properties
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Appearance
Solution
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Formulation
Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.
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Concentration
Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration
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Storage & Stability
Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.
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Shipping
Shipping with blue ice.
Verification Images
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Western blot analysis of extracts from A431 (lane 2(20μg) and A431 (lane 3(40μg)using ASK1 (HY-P80020) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% defatted milk powder in TBST for 2 hour at room temperature. The primary antibody (HY-P80020, 1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% defatted milk powder in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
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Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue using ASK1 Antibody (YA609). The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80020, 1/100) in 4℃ overnight. 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. -
Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue using ASK1 Antibody (YA609). The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80020, 1/100) in 4℃ overnight. 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. -
Flow cytometric analysis of 1X106 HeLa cells labeling ASK1 Antibody(HY-P80020, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/100 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).
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Immunocytochemistry analysis of Hela cells labeling ASK1 with ASK1 Antibody (YA609) (HY-P80020) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with ASK1 Antibody (YA609) (HY-P80020) at 1/500 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L(HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
Background
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Function
Apoptosis signal-regulating kinase 1 (ASK1) is a member of the mitogen-activated protein kinase (MAP3K) family which acts as an upstream regulator for the activation of p38 MAPK and c-Jun N-terminal kinase (JNK) . Under normal physiological conditions, ASK1 is maintained in an inactive state through binding to the reduced form of thioredoxin. However, during pathological conditions which induce intracellular production of reactive oxygen species (e.g., mitochondrial or endoplasmic reticulum stress), thioredoxin is oxidized and dissociates from ASK1 facilitating activation of ASK1 and downstream p38 MAPK and JNK signaling that can cause tissue injury .
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Subcellular Localization
Cytoplasm; Endoplasmic reticulum
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Expression
Tissue_specificity:It is expressed in large quantities in the heart and pancreas.
Induction:By TNF. Inhibited by HIV-1 Nef -
Subunit
Homodimer when inactive. Binds both upstream activators and downstream substrates in multimolecular complexes. Part of a cytoplasmic complex made of HIPK1, DAB2IP and MAP3K5 in response to TNF (PubMed:15310755, PubMed:15701637, PubMed:17210579, PubMed:17389591). This complex formation promotes MAP3K5-JNK activation and subsequent apoptosis. Interacts with SOCS1 which recognizes phosphorylation of Tyr-718 and induces MAP3K5/ASK1 degradation in endothelial cells. Interacts with the 14-3-3 family proteins such as YWHAB, YWHAE, YWHAQ, YWHAH, YWHAZ and SFN (PubMed:10411906, PubMed:15023544, PubMed:15094778). Interacts with ARRB2, BIRC2, DAB2IP, IGF1R, MAP3K6/ASK2, PGAM5, PIM1, PPP5C, SOCS1, STUB1, TRAF2, TRAF6 and TXN (PubMed:10688666, PubMed:11090355, PubMed:11689443, PubMed:12556535, PubMed:12813029, PubMed:15310755, PubMed:16038411, PubMed:16129676, PubMed:16407264, PubMed:17220297, PubMed:17724081, PubMed:18408005, PubMed:19590015, PubMed:19749799, PubMed:9564042, PubMed:9774977). Interacts with ERN1 in a TRAF2-dependent manner (PubMed:14749717). Interacts with calcineurin subunit PPP3R1 (By similarity). Interacts with PPM1L (PubMed:17456047). Interacts (via N-terminus) with RAF1 and this interaction inhibits the proapoptotic function of MAP3K5 (PubMed:11427728). Interacts with DAB2IP (via N-terminus C2 domain); the interaction occurs in a TNF-alpha-dependent manner (PubMed:15310755). Interacts with DUSP13A; may positively regulate apoptosis (PubMed:20358250). Interacts with DAXX (PubMed:9743501). Interacts with RC3H2 (PubMed:24448648). Interacts with PPIA/CYPA (PubMed:26095851). Interacts with PRMT1; the interaction results in MAP3K5 methylation by PRMT1 which inhibits MAP3K5 activation (PubMed:22095282). Interacts with TRAF2; the interaction is inhibited by PRMT1 (PubMed:22095282). Interacts with TRIM48 (PubMed:29186683)
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SwissProt ID
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Synonyms
ASK1, MAPKKK5, MEKK5, MAP3K5, Mitogen-activated protein kinase kinase kinase 5, Apoptosis signal-regulating kinase 1, MAPK/ERK kinase kinase 5, ASK-1, MEK kinase 5, MEKK 5
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Research Field
Cell Biology
Documentation
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Data Sheet (235 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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User Guide for Antibodies (1077 KB)