PRODUCT CODE: ET1609-42

Recombinant Phospho JNK1/2/3(T183+T183+T221) Monoclonal Antibody (ET1609-42)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of Phospho-JNK1/2/3(T183+T183+T221) on different lysates using anti-Phospho-JNK1/2/3(T183+T183+T221) antibody at 1/500 dilution.<br />
Lane 1: NIH/3T3 cell lysate, treated with Anisomycin<br />
Lane 2: NIH/3T3 cell lysate, untreated
  • Western blot analysis of Phospho-JNK1/2/3(T183+T183+T221) on different lysates using anti-Phospho-JNK1/2/3(T183+T183+T221) antibody at 1/500 dilution.<br />
Lane 1: NIH/3T3 cell lysate, treated with Anisomycin<br />
Lane 2: NIH/3T3 cell lysate, untreated
  • ICC staining of Phospho-JNK1/2/3(T183+T183+T221) 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 (ET1609-42, 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).
  • ICC staining of Phospho-JNK1/2/3(T183+T183+T221) in NIH/3T3 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 (ET1609-42, 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).
  • ICC staining of Phospho-JNK1/2/3(T183+T183+T221) in HUVEC 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 (ET1609-42, 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).
  • Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-Phospho-JNK1/2/3(T183+T183+T221) 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 (ET1609-42, 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.
  • Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-Phospho-JNK1/2/3(T183+T183+T221) 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 (ET1609-42, 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.
  • Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-Phospho-JNK1/2/3(T183+T183+T221) 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 (ET1609-42, 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.
  • Flow cytometric analysis of Phospho-JNK1/2/3(T183+T183+T221) was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1609-42, 1/100) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
Western blot analysis of Phospho-JNK1/2/3(T183+T183+T221) on different lysates using anti-Phospho-JNK1/2/3(T183+T183+T221) antibody at 1/500 dilution.
Lane 1: NIH/3T3 cell lysate, treated with Anisomycin
Lane 2: NIH/3T3 cell lysate, untreated

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant Phospho JNK1/2/3(T183+T183+T221) Monoclonal Antibody (ET1609-42)

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding thr183 + thr183 + thr221 of human jnk1/2/3

Host

Rabbit

Modification

Phospho

Modification Site

JNK1(T183)+JNK2(T183)+JNK3(T221)

Positive Control

HUVEC, NIH/3T3, Hela, mouse heart tissue, human liver tissue, human uterus tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

ST500

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

54/46 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

Mitogen-activated protein kinase 8

GENE NAME

MAPK8

SYNONYMS

MAP kinase 8/9/10, MAPK 8/9/10, SAPK1c, SAPK1a, SAPK1b, MAPK8/9/10, JNK1, PRKM8, SAPK1, SAPK1C, JNK2, PRKM9, SAPK1A, JNK3, JNK3A, PRKM10, SAPK1B

SEQUENCE SIMILARITIES

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

POST-TRANSLATIONAL MODIFICATION

Dually phosphorylated on Thr-183 and Tyr-185 by MAP2K7 and MAP2K4, which activates the enzyme. Phosphorylated by TAOK2. May be phosphorylated at Thr-183 and Tyr-185 by MAP3K1/MEKK1. Phosphorylated form is more concentrated at synapses than none-phosphorylated (By similarity).

SUBCELLULAR LOCATION

Cytoplasm. Nucleus. Cell junction, synapse. Note=In the cortical neurons, predominantly cytoplasmic and associated with the Golgi apparatus and endosomal fraction. Increased neuronal activity increases phosphorylated form at synapses (By similarity). Colocalizes with POU5F1 in the nucleus.

FUNCTION

Serine/threonine-protein kinase involved in various processes such as cell proliferation, differentiation, migration, transformation and programmed cell death. Extracellular stimuli such as proinflammatory cytokines or physical stress stimulate the stress-activated protein kinase/c-Jun N-terminal kinase (SAP/JNK) signaling pathway. In this cascade, two dual specificity kinases MAP2K4/MKK4 and MAP2K7/MKK7 phosphorylate and activate MAPK8/JNK1. In turn, MAPK8/JNK1 phosphorylates a number of transcription factors, primarily components of AP-1 such as JUN, JDP2 and ATF2 and thus regulates AP-1 transcriptional activity. Phosphorylates the replication licensing factor CDT1, inhibiting the interaction between CDT1 and the histone H4 acetylase HBO1 to replication origins. Loss of this interaction abrogates the acetylation required for replication initiation. Promotes stressed cell apoptosis by phosphorylating key regulatory factors including p53/TP53 and Yes-associates protein YAP1. In T-cells, MAPK8 and MAPK9 are required for polarized differentiation of T-helper cells into Th1 cells. Contributes to the survival of erythroid cells by phosphorylating the antagonist of cell death BAD upon EPO stimulation. Mediates starvation-induced BCL2 phosphorylation, BCL2 dissociation from BECN1, and thus activation of autophagy. Phosphorylates STMN2 and hence regulates microtubule dynamics, controlling neurite elongation in cortical neurons. In the developing brain, through its cytoplasmic activity on STMN2, negatively regulates the rate of exit from multipolar stage and of radial migration from the ventricular zone. Phosphorylates several other substrates including heat shock factor protein 4 (HSF4), the deacetylase SIRT1, ELK1, or the E3 ligase ITCH. Phosphorylates the CLOCK-ARNTL/BMAL1 heterodimer and plays a role in the regulation of the circadian clock. Phosphorylates the heat shock transcription factor HSF1, suppressing HSF1-induced transcriptional activity. Phosphorylates POU5F1, which results in the inhibition of POU5F1's transcriptional activity and enhances its proteosomal degradation (By similarity). Phosphorylates JUND and this phosphorylation is inhibited in the presence of MEN1 (By similarity). In neurons, phosphorylates SYT4 which captures neuronal dense core vesicles at synapses (By similarity).; JNK1 isoforms display different binding patterns: beta-1 preferentially binds to c-Jun, whereas alpha-1, alpha-2, and beta-2 have a similar low level of binding to both c-Jun or ATF2. However, there is no correlation between binding and phosphorylation, which is achieved at about the same efficiency by all isoforms.