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  • Product Name
    Phospho-ASK1 (Ser966) Antibody
  • Catalog No.
    AF3477
  • RRID
    AB_2834915
  • Source
    Rabbit
  • Application
    WB,IHC,IF/ICC,ELISA
  • Reactivity
    Human, Mouse, Rat
  • Prediction
    Pig(100%), Zebrafish(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%)
  • UniProt
  • Mol.Wt
    (Observed)155kD; (Calculated)155kDa
  • Concentration
    1mg/ml
  • Browse similar products>>

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Product Information

Alternative Names:Expand▼

Apoptosis signal regulating kinase 1; Apoptosis signal-regulating kinase 1; ASK 1; ASK-1; ASK1; M3K5; M3K5_HUMAN; MAP/ERK kinase kinase 5; MAP3K5; MAPK/ERK kinase kinase 5; MAPKKK5; MEK kinase 5; MEKK 5; MEKK5; Mitogen activated protein kinase kinase kinase 5; Mitogen-activated protein kinase kinase kinase 5;

Applications:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000

Reactivity:

Human, Mouse, Rat

Predicted Reactivity:

Pig(100%), Zebrafish(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%)

Source:

Rabbit

Clonality:

Polyclonal

Purification:

The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.

Specificity:

Phospho-ASK1 (Ser966) Antibody detects endogenous levels of ASK1 only when phosphorylated at Serine 966.

RRID:

AB_2834915
Please cite this product as: Affinity Biosciences Cat# AF3477, RRID:AB_2834915.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Store at -20 °C.Stable for 12 months from date of receipt.

Immunogen Information

Immunogen:

A synthesized peptide derived from human ASK1 around the phosphorylation site of Ser966.

Uniprot:



>>Visit The Human Protein Atlas

Gene ID:

Gene Name:

MAP3K5

Molecular Weight:

Observed Mol.Wt.: (Observed)155kD.
Predicted Mol.Wt.: (Calculated)155kDa.

Tissue Specificity:

Q99683 M3K5_HUMAN:
Abundantly expressed in heart and pancreas.

Description:

Mitogen-activated protein kinase (MAPK) signaling cascades include MAPK or extracellular signal-regulated kinase (ERK), MAPK kinase (MKK or MEK), and MAPK kinase kinase (MAPKKK or MEKK). MAPKK kinase/MEKK phosphorylates and activates its downstream protein kinase, MAPK kinase/MEK, which in turn activates MAPK.

Sequence:
MSTEADEGITFSVPPFAPSGFCTIPEGGICRRGGAAAVGEGEEHQLPPPPPGSFWNVESAAAPGIGCPAATSSSSATRGRGSSVGGGSRRTTVAYVINEASQGQLVVAESEALQSLREACETVGATLETLHFGKLDFGETTVLDRFYNADIAVVEMSDAFRQPSLFYHLGVRESFSMANNIILYCDTNSDSLQSLKEIICQKNTMCTGNYTFVPYMITPHNKVYCCDSSFMKGLTELMQPNFELLLGPICLPLVDRFIQLLKVAQASSSQYFRESILNDIRKARNLYTGKELAAELARIRQRVDNIEVLTADIVINLLLSYRDIQDYDSIVKLVETLEKLPTFDLASHHHVKFHYAFALNRRNLPGDRAKALDIMIPMVQSEGQVASDMYCLVGRIYKDMFLDSNFTDTESRDHGASWFKKAFESEPTLQSGINYAVLLLAAGHQFESSFELRKVGVKLSSLLGKKGNLEKLQSYWEVGFFLGASVLANDHMRVIQASEKLFKLKTPAWYLKSIVETILIYKHFVKLTTEQPVAKQELVDFWMDFLVEATKTDVTVVRFPVLILEPTKIYQPSYLSINNEVEEKTISIWHVLPDDKKGIHEWNFSASSVRGVSISKFEERCCFLYVLHNSDDFQIYFCTELHCKKFFEMVNTITEEKGRSTEEGDCESDLLEYDYEYDENGDRVVLGKGTYGIVYAGRDLSNQVRIAIKEIPERDSRYSQPLHEEIALHKHLKHKNIVQYLGSFSENGFIKIFMEQVPGGSLSALLRSKWGPLKDNEQTIGFYTKQILEGLKYLHDNQIVHRDIKGDNVLINTYSGVLKISDFGTSKRLAGINPCTETFTGTLQYMAPEIIDKGPRGYGKAADIWSLGCTIIEMATGKPPFYELGEPQAAMFKVGMFKVHPEIPESMSAEAKAFILKCFEPDPDKRACANDLLVDEFLKVSSKKKKTQPKLSALSAGSNEYLRSISLPVPVLVEDTSSSSEYGSVSPDTELKVDPFSFKTRAKSCGERDVKGIRTLFLGIPDENFEDHSAPPSPEEKDSGFFMLRKDSERRATLHRILTEDQDKIVRNLMESLAQGAEEPKLKWEHITTLIASLREFVRSTDRKIIATTLSKLKLELDFDSHGISQVQVVLFGFQDAVNKVLRNHNIKPHWMFALDSIIRKAVQTAITILVPELRPHFSLASESDTADQEDLDVEDDHEEQPSNQTVRRPQAVIEDAVATSGVSTLSSTVSHDSQSAHRSLNVQLGRMKIETNRLLEELVRKEKELQALLHRAIEEKDQEIKHLKLKSQPIEIPELPVFHLNSSGTNTEDSELTDWLRVNGADEDTISRFLAEDYTLLDVLYYVTRDDLKCLRLRGGMLCTLWKAIIDFRNKQT

Research Background

Function:

Serine/threonine kinase which acts as an essential component of the MAP kinase signal transduction pathway. Plays an important role in the cascades of cellular responses evoked by changes in the environment. Mediates signaling for determination of cell fate such as differentiation and survival. Plays a crucial role in the apoptosis signal transduction pathway through mitochondria-dependent caspase activation. MAP3K5/ASK1 is required for the innate immune response, which is essential for host defense against a wide range of pathogens. Mediates signal transduction of various stressors like oxidative stress as well as by receptor-mediated inflammatory signals, such as the tumor necrosis factor (TNF) or lipopolysaccharide (LPS). Once activated, acts as an upstream activator of the MKK/JNK signal transduction cascade and the p38 MAPK signal transduction cascade through the phosphorylation and activation of several MAP kinase kinases like MAP2K4/SEK1, MAP2K3/MKK3, MAP2K6/MKK6 and MAP2K7/MKK7. These MAP2Ks in turn activate p38 MAPKs and c-jun N-terminal kinases (JNKs). Both p38 MAPK and JNKs control the transcription factors activator protein-1 (AP-1).

Post-translational Modifications:

Phosphorylated at Thr-838 through autophosphorylation and by MAP3K6/ASK2 which leads to activation. Thr-838 is dephosphorylated by PPP5C. Ser-83 and Ser-1033 are inactivating phosphorylation sites, the former of which is phosphorylated by AKT1 and AKT2. Phosphorylated at Ser-966 which induces association of MAP3K5/ASK1 with the 14-3-3 family proteins and suppresses MAP3K5/ASK1 activity. Calcineurin (CN) dephosphorylates this site. Also dephosphorylated and activated by PGAM5.

Ubiquitinated. Tumor necrosis factor (TNF) induces TNFR2-dependent ubiquitination leading to proteasomal degradation.

Subcellular Location:

Cytoplasm. Endoplasmic reticulum.
Note: Interaction with 14-3-3 proteins alters the distribution of MAP3K5/ASK1 and restricts it to the perinuclear endoplasmic reticulum region.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionGraphics by Christian Stolte

Tissue Specificity:

Abundantly expressed in heart and pancreas.

Subunit Structure:

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:9564042,PubMed:9774977, 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). Interacts with ERN1 in a TRAF2-dependent manner (PubMed:14749717). Interacts with calcineurin subunit PPP3R1 and with PPM1L (PubMed:17456047) (By similarity). 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 DUSP13/DUSP13A; may positively regulate apoptosis (PubMed:20358250). Interacts with DAXX (PubMed:9743501). Interacts with RC3H2 (PubMed:24448648).

(Microbial infection) Interacts with HIV-1 Nef; this interaction inhibits MAP3K5 signaling.

Similarity:

Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.

Research Fields

Research Fields:

· Cellular Processes > Cellular community - eukaryotes > Tight junction.(View pathway)
· Cellular Processes > Cell growth and death > Apoptosis.(View pathway)
· Environmental Information Processing > Signal transduction > TNF signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > MAPK signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Sphingolipid signaling pathway.(View pathway)
· Genetic Information Processing > Folding, sorting and degradation > Protein processing in endoplasmic reticulum.(View pathway)
· Human Diseases > Neurodegenerative diseases > Amyotrophic lateral sclerosis (ALS).
· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.
· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).
· Organismal Systems > Nervous system > Neurotrophin signaling pathway.(View pathway)

WB Images(1)


IHC Images(1)


IF/ICC Images(1)


ELISA analysis of AF3477 showing specificity to Phospho-ASK1 (Ser966) Blocking Peptide peptide. Peptides concentration: 1ug/ml.
P-peptide: phospho-peptide; N-peptide: non-phospho-peptide.

Reference Citations:

1). Chenxi Shi et al. JNK signaling pathway mediates acetaminophen-induced hepatotoxicity accompanied by changes of glutathione S-transferase A1 content and expression. FRONT PHARMACOL 2019 Sep [IF=3.845]

2). Shi C et al. JNK Signaling Pathway Mediates Acetaminophen-Induced Hepatotoxicity Accompanied by Changes of Glutathione S-Transferase A1 Content and Expression. Front Pharmacol 2019 Sep 20;10:1092 (PubMed: 31620005) [IF=3.845]

3). Lu M et al. CTGF Triggers Rat Astrocyte Activation and Astrocyte-Mediated Inflammatory Response in Culture Conditions. Inflammation 2019 Jun 10 (PubMed: 31183597)

4). Feng H et al. Dual function of peroxiredoxin I in lipopolysaccharide-induced osteoblast apoptosis via reactive oxygen species and the apoptosis signal-regulating kinase 1 signaling pathway. Cell Death Discov 2018 Apr 27;4:47 (PubMed: 29707240)

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Catalog Number :

AF3477-BP
(Blocking peptide available as AF3477-BP)

Price/Size :

$350/1mg.
Tips: For phospho antibody, we provide phospho peptide(0.5mg) and non-phospho peptide(0.5mg).

Function :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Zebrafish
100%
Chicken
100%
Rabbit
100%
Pig
100%
Dog
100%
Horse
100%
Bovine
100%
Sheep
100%
Xenopus
0%
High similarity Medium similarity Low similarity No similarity
Q99683 as Substrate
Site PTM Type Enzyme
S82 Phosphorylation
S83 Phosphorylation P11309 (PIM1) , P51812 (RPS6KA3) , P31749 (AKT1) , P31751 (AKT2)
R89 Methylation
S101 Phosphorylation
S115 Phosphorylation
S461 Phosphorylation
K465 Ubiquitination
T528 Phosphorylation
K535 Sumoylation
Y570 Phosphorylation
Y574 Phosphorylation
K709 Ubiquitination
Y718 Phosphorylation P23458 (JAK1) , O60674 (JAK2)
K730 Ubiquitination
K785 Ubiquitination
T813 Phosphorylation Q99683 (MAP3K5)
Y814 Phosphorylation
S821 Phosphorylation
T825 Phosphorylation
K827 Ubiquitination
T838 Phosphorylation Q14680 (MELK) , Q99683 (MAP3K5) , O95382 (MAP3K6) , Q15139 (PRKD1)
T842 Phosphorylation Q99683 (MAP3K5)
C869 S-Nitrosylation
K946 Acetylation
S958 Phosphorylation
S964 Phosphorylation
S966 Phosphorylation O15530 (PDPK1) , Q99683 (MAP3K5)
Y982 Phosphorylation
S986 Phosphorylation
S997 Phosphorylation
S1029 Phosphorylation
S1033 Phosphorylation
T1059 Phosphorylation
K1064 Ubiquitination
K1083 Sumoylation
T1109 Phosphorylation P51812 (RPS6KA3)
K1114 Sumoylation
S1240 Phosphorylation
T1326 Phosphorylation P51812 (RPS6KA3)
Q99683 as PTM Enzyme
Substrate Site Source
O15530 (PDPK1) S394 Uniprot
O15530 (PDPK1) S398 Uniprot
P38936 (CDKN1A) S98 Uniprot
P45379-11 (TNNT2) T191 Uniprot
P45379-6 (TNNT2) T194 Uniprot
P45379-11 (TNNT2) S195 Uniprot
P45379-6 (TNNT2) S198 Uniprot
P45379-10 (TNNT2) T201 Uniprot
P45379 (TNNT2) T204 Uniprot
P45379-10 (TNNT2) S205 Uniprot
P45379 (TNNT2) S208 Uniprot
P45983 (MAPK8) T183 Uniprot
P45983 (MAPK8) Y185 Uniprot
P45985 (MAP2K4) T261 Uniprot
P46734 (MAP2K3) S218 Uniprot
P52564 (MAP2K6) S207 Uniprot
Q16539 (MAPK14) T180 Uniprot
Q16539 (MAPK14) Y182 Uniprot
Q969S3 (ZNF622) S314 Uniprot
Q969S3 (ZNF622) T318 Uniprot
Q99683 (MAP3K5) T813 Uniprot
Q99683 (MAP3K5) T838 Uniprot
Q99683 (MAP3K5) T842 Uniprot
Q99683 (MAP3K5) S966 Uniprot
Q9UER7 (DAXX) S176 Uniprot
Q9UER7 (DAXX) S184 Uniprot
Q9Y3F4 (STRAP) T175 Uniprot
Q9Y3F4 (STRAP) S179 Uniprot
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.

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