Human/Mouse/Rat Phospho-JNK (T183/Y185) Antibody

R&D Systems | Catalog # AF1205

R&D Systems
100% Guarantee

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Simple Western

Cited:

Immunohistochemistry, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
View all JNK Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing human, rat, and mouse JNK1 T183/Y185 site

Specificity

Detects human, mouse and rat p46 and p54 JNK when dually phosphorylated at sites homologous to T183/Y185 of JNK1 and JNK2, and T221/Y223 of JNK3 in Western blots.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat Phospho-JNK (T183/Y185) Antibody

Detection of Human and Mouse Phospho-JNK (T183/Y185) antibody by Western Blot.

Detection of Human and Mouse Phospho-JNK (T183/Y185) by Western Blot.

Western blot shows lysates of HEK293 human embryonic kidney cell line and NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 J/m2UV-C for 30 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-JNK (T183/Y185) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1205), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for Phospho-JNK (T183/Y185) at approximately 46 and 54 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 6.
JNK antibody in Human Breast Cancer Tissue by Immunohistochemistry (IHC-P).

JNK in Human Breast Cancer Tissue.

JNK phosphorylated at T183/Y185 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human/Mouse/Rat Phospho-JNK (T183/Y185) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1205) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of immersion fixed paraffin-embedded Tissue Sections.
JNK antibody in Human Prostate by Immunohistochemistry (IHC-P).

JNK in Human Prostate.

JNK phosphorylated at T183/Y185 was detected in immersion fixed paraffin-embedded sections of human prostate array using Rabbit Anti-Human/Mouse/Rat Phospho-JNK (T183/Y185) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1205) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Phospho-JNK (T183/Y185) antibody by Simple WesternTM.

Detection of Human Phospho-JNK (T183/Y185) by Simple WesternTM.

Simple Western lane view shows lysates of HEK293T human embryonic kidney cell line untreated (-) or treated (+) with 100 J/m2ultraviolet light (UV) for 30 minutes, loaded at 0.2 mg/mL. Specific bands were detected for Phospho-JNK (T183/Y185) at approximately 56 and 46 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-JNK (T183/Y185) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1205). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.*Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
Detection of Mouse JNK1/2/3 by Western Blot

Detection of Mouse JNK1/2/3 by Western Blot

Effect of p38MAPK (p38) inhibition on intracellular signaling following IR. Rats were pretreated with the carrier DMSO or BIRB796 (B-796) (5 mg/kg BW) for 1 hour and subjected to 1 hour of renal ischemia followed by different time points of reperfusion (15 min, 2 days, 7 days). Kidneys were harvested at given time points of reperfusion and total tissue lysates were used to determine activation pattern of MAPKs (p38MAPK, JNK, ERK) and the downstream target of p38MAPK (MK2) by phosphorylation specific antibodies. A representative immunoblot (A) and summary graphs (B, C) are shown. Results are given as mean ± SEM (n = 3). ** p < 0.01, ***p < 0.001 vs. sham-operated group; §p < 0.01, #p < 0.001 vs. IR-15 min group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24423080), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse JNK1/2/3 by Western Blot

Detection of Mouse JNK1/2/3 by Western Blot

Inhibition of JNK kinases prevents p66ShcS36 phosphorylation under oxidative stress.MEFs were treated with H2O2 (a) (1 mM, 30 min) or exposed to HR (hypoxia 90 min, reoxygenation 15 min) (b). Cell lysates were probed with the antibodies indicated to assess p66Shc phosphorylation at S36. Representative Western blots (individual experiment performed under the same experimental conditions and run on the same gel) and summary graphs from at least three independent experiments are shown. Signal intensity of control samples was arbitrarily set at 1. Mitochondrial ROS production was measured in p66Shc+/+ and p66Shc−/− MEFs after prooxidant treatment as described in Material and Methods (n ≥ 4) (c) and phospho gamma H2AX was detected by immunoblotting (d, lower panel) and summary graph is shown in above panel. SP: SP600125, JNK inhibitor, and BR: BR796, p38 inhibitor were applied 1 h prior to stress application. Original blot shown in (b) have been cropped and the full length blots are shown in the Supplementary Figure S1b Statistical analysis was performed using t-test or ANOVA (*p < 0.05 **p < 0.01, ***p < 0.001). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26868434), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse JNK1/2/3 by Western Blot

Detection of Mouse JNK1/2/3 by Western Blot

The impact of high glucose conditions on various down-stream signaling pathways.The status of various signaling pathways in retinal AC under various glucose conditions was determined as described in Methods. Levels of active Src (A), Akt (B), ERK (C), p38 (D), JNK1 (E), Fyn (F) and p65 (G) in retinal AC under different glucose conditions were determined by Western blot analysis from equal amounts of cell lysates. The total level for each protein is shown in the lower part of each panel. The quantitative assessment of the data for each blot is shown below the blot. Data are presented as mean ± SEM. n = 3; *p<0.05 (NG vs. HG and NG vs. NG+L-Glu). Levels of active Fyn (F) in retinal AC under different glucose conditions was determined by Western blot analysis of immunoprecipitates from equal amounts of retinal AC lysates. Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0103148), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse JNK1/2/3 by Western Blot

Detection of Mouse JNK1/2/3 by Western Blot

Effect of p38MAPK (p38) inhibition on intracellular signaling following IR. Rats were pretreated with the carrier DMSO or BIRB796 (B-796) (5 mg/kg BW) for 1 hour and subjected to 1 hour of renal ischemia followed by different time points of reperfusion (15 min, 2 days, 7 days). Kidneys were harvested at given time points of reperfusion and total tissue lysates were used to determine activation pattern of MAPKs (p38MAPK, JNK, ERK) and the downstream target of p38MAPK (MK2) by phosphorylation specific antibodies. A representative immunoblot (A) and summary graphs (B, C) are shown. Results are given as mean ± SEM (n = 3). ** p < 0.01, ***p < 0.001 vs. sham-operated group; §p < 0.01, #p < 0.001 vs. IR-15 min group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24423080), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse JNK1/2/3 by Western Blot

Detection of Mouse JNK1/2/3 by Western Blot

Inhibition of JNK kinases prevents p66ShcS36 phosphorylation under oxidative stress.MEFs were treated with H2O2 (a) (1 mM, 30 min) or exposed to HR (hypoxia 90 min, reoxygenation 15 min) (b). Cell lysates were probed with the antibodies indicated to assess p66Shc phosphorylation at S36. Representative Western blots (individual experiment performed under the same experimental conditions and run on the same gel) and summary graphs from at least three independent experiments are shown. Signal intensity of control samples was arbitrarily set at 1. Mitochondrial ROS production was measured in p66Shc+/+ and p66Shc−/− MEFs after prooxidant treatment as described in Material and Methods (n ≥ 4) (c) and phospho gamma H2AX was detected by immunoblotting (d, lower panel) and summary graph is shown in above panel. SP: SP600125, JNK inhibitor, and BR: BR796, p38 inhibitor were applied 1 h prior to stress application. Original blot shown in (b) have been cropped and the full length blots are shown in the Supplementary Figure S1b Statistical analysis was performed using t-test or ANOVA (*p < 0.05 **p < 0.01, ***p < 0.001). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26868434), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Phospho-JNK (T183/Y185) by Western Blot

Detection of Phospho-JNK (T183/Y185) by Western Blot

Activation of ERK1/2 signaling was necessary for venlafaxine-induced apoptosis. (A) Representative western-blot bands and quantification of the MAPK signal molecules abundances in MV3 cells treated with venlafaxine (10 μM) for 0–8 h (B) MV3 cells were treated with vehicle, JNK1/2 inhibitor SP600125 and MERK/ERK inhibitor PD98059 for 72 h. Cell viability were measured by CCK-8 kits at 72 h (C) MV3 cells were transfected with siRNA control, JNK1/2 siRNA, and ERK siRNA for 72 h. Cell viability were measured by CCK-8 kits at 72 h (D,E) Representative western-blot bands and quantification of Nur77 abundances in MV3 cells treated with venlafaxine (10 μM) for 8 h, or transfected with siRNA control, JNK1/2 siRNA, and ERK siRNA for 20 h (F) MV3 cells were immunostained with Bcl-2 and Nur77 antibodies and visualized by confocal microscopy. N = 3, *, p <.05, **, <.01, ***, p <.001 vs. control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36686679), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Phospho-JNK (T183/Y185) by Western Blot

Detection of Phospho-JNK (T183/Y185) by Western Blot

Activation of ERK1/2 signaling was necessary for venlafaxine-induced apoptosis. (A) Representative western-blot bands and quantification of the MAPK signal molecules abundances in MV3 cells treated with venlafaxine (10 μM) for 0–8 h (B) MV3 cells were treated with vehicle, JNK1/2 inhibitor SP600125 and MERK/ERK inhibitor PD98059 for 72 h. Cell viability were measured by CCK-8 kits at 72 h (C) MV3 cells were transfected with siRNA control, JNK1/2 siRNA, and ERK siRNA for 72 h. Cell viability were measured by CCK-8 kits at 72 h (D,E) Representative western-blot bands and quantification of Nur77 abundances in MV3 cells treated with venlafaxine (10 μM) for 8 h, or transfected with siRNA control, JNK1/2 siRNA, and ERK siRNA for 20 h (F) MV3 cells were immunostained with Bcl-2 and Nur77 antibodies and visualized by confocal microscopy. N = 3, *, p <.05, **, <.01, ***, p <.001 vs. control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36686679), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Rat Phospho-JNK (T183/Y185) by Western Blot

Detection of Rat Phospho-JNK (T183/Y185) by Western Blot

Evaluation of the phosphorylation of MAPK family members and Raf-1 in Bag-1-overexpressing NP cells. Western blot analysis (A) and quantification (B,C) of the phosphorylated forms of p38, ERK, JNK, and Raf-1 in NP cells treated with or without H2O2 for 30 min. Data are the means ± SD n = 3, * p < 0.05. (D) Comparison of the expression of pERK in NP cells (NP) and Bag-1-overexpressing NP cells (Bag) without H2O2 treatment. Data are the means ± SD n = 3, * p < 0.05. Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/2227-9059/11/3/863), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse/Rat Phospho-JNK (T183/Y185) Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human prostate cancer tissue and human breast cancer tissue

Simple Western

5 µg/mL
Sample: HEK293T human embryonic kidney cell line treated with ultraviolet light (UV)

Western Blot

0.5 µg/mL
Sample: UV-C-treated HEK293 human embryonic kidney cell line and NIH‑3T3 mouse embryonic fibroblast cell line

Reviewed Applications

Read 4 reviews rated 3.5 using AF1205 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen and protein A Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

=
÷

Background: JNK

The c-Jun N-terminal Kinases (JNKs) are part of the MAPK (mitogen-activated protein kinase) system that transmits signals from the extracellular milieu to both the cytoplasm and nucleus of the cell. Following perturbation at the cell membrane, MEKKs/MAP3Ks are initially activated, followed by their activation of MKKs/MAP2Ks, and MKKs activation of MAPKs/MAP(1)Ks. There are three classes of MAPKs: ERKs, p38 Kinases and JNKs. JNKs are 45-55 kDa protein products of three genes which, through alternative splicing, generate up to 10 possible isoforms. The phosphorylation targets for MAPKs vary, but include p53, c-MYC, ATF2 and c-Jun, the latter molecule representing the namesake for the enzyme group. The three human JNKs share approximately 80% aa sequence identity. JNKs from human, mouse and rat all contain a conserved Met-Met-Thr(183)-Pro-Tyr(185)-Val-Val motif that undergoes dual phosphorylation by MMK4 and MMK7 to activate the different JNKs. Activated by environmental stresses and inflammatory cytokines, JNKs translocate to the nucleus where they regulate the activity of several transcription factors; including the c-Jun component of AP-1 and ATF-2.

Long Name

C-Jun N-terminal Kinase

Alternate Names

c-Jun N-terminal kinase 1, EC 2.7.11, EC 2.7.11.24, JNK1 alpha protein kinase, JNK1 beta protein kinase, JNK1JNK1A2, JNK-46, JUN N-terminal kinase, MAP kinase 8, MAPK 8, mitogen-activated protein kinase 8, mitogen-activated protein kinase 8 isoform JNK1 alpha1, mitogen-activated protein kinase 8 isoform JNK1 beta2, PRKM8JNK, protein kinase JNK1, SAPK1JNK21B1/2, Stress-activated protein kinase 1, Stress-activated protein kinase JNK1

Additional JNK Products

Product Documents for Human/Mouse/Rat Phospho-JNK (T183/Y185) Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human/Mouse/Rat Phospho-JNK (T183/Y185) Antibody

For research use only

Citations for Human/Mouse/Rat Phospho-JNK (T183/Y185) Antibody

Customer Reviews for Human/Mouse/Rat Phospho-JNK (T183/Y185) Antibody (4)

3.5 out of 5
4 Customer Ratings
5 Stars
0%
4 Stars
50%
3 Stars
50%
2 Stars
0%
1 Stars
0%

Have you used Human/Mouse/Rat Phospho-JNK (T183/Y185) Antibody?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Customer Images

Showing  1 - 4 of 4 reviews Showing All
Filter By:
Clear Filters X
  • Human/Mouse/Rat Phospho-JNK (T183/Y185) Antibody
    Name: Anonymous
    Application: Microarrays
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 01/14/2021
    Human/Mouse/Rat Phospho-JNK (T183/Y185) Antibody AF1205
  • Human/Mouse/Rat Phospho-JNK (T183/Y185) Antibody
    Name: Anonymous
    Application: Microarrays
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 11/14/2018
  • Human/Mouse/Rat Phospho-JNK (T183/Y185) Antibody
    Name: Carly Garrison
    Application: Microarray
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 10/31/2018
  • Human/Mouse/Rat Phospho-JNK (T183/Y185) Antibody
    Name: Aleksandra Hac
    Application: Western Blot
    Sample Tested: MEF whole cell lysate, PC3 Whole Cell Lysate (Human prostate cancer cell) and HDFa whole cell lysate
    Species: Human and Mouse
    Verified Customer | Posted 05/04/2017
    We obtained bands at predicted molecular weight for both human (PC3, HDFa) as well as mouse (MEF) samples. However, at recommended concentrations the signal was quite weak using ECL substrate. Perhaps they work better using substrates with higher sensitivity.

There are no reviews that match your criteria.

Showing  1 - 4 of 4 reviews Showing All

Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs

No product specific FAQs exist for this product.

View all FAQs for Antibodies