Name: Anti-c-Jun (phospho S63) antibody [Y172]
Brand: RabMAb
SKU: ab32385
Rating: 5 (4 reviews)

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Immunocytochemistry/ Immunofluorescence - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [Y172] to c-Jun (phospho S63)
Suitable for: WB, IHC-P, ICC/IF, Dot blot, ELISA
Reacts with: Mouse, Human

Carrier Free

Product name

Anti-c-Jun (phospho S63) antibody [Y172]
See all c-Jun primary antibodies
Description

Rabbit monoclonal [Y172] to c-Jun (phospho S63)
Host species

Rabbit
Specificity

Anti-c-Jun (phospho S63) antibody [Y172] (ab32385) only detects c-Jun phosphorylated on Serine 63 when tested in WB and ICC using specific phospho-treatments. However, in DotBlot and ELISA assays we detected some cross-reactivity with the non-phospho peptide as well. Please refer to the images on the datasheet. The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.
Tested applications

Suitable for: WB, IHC-P, ICC/IF, Dot blot, ELISAmore details
Unsuitable for: Flow Cyt
Species reactivity

Reacts with: Mouse, Human
Predicted to work with: Rat, Cow
Immunogen

Synthetic peptide within Human c-Jun aa 50-150 (phospho S63). The exact sequence is proprietary.
Database link: P05412
Positive control
- WB: UV or Anisomycin treated NIH/3T3 or HeLa whole cell lysate (ab150035). IHC-P: Human breast carcinoma tissue. ICC/IF: A431 cells, NIH/3T3 cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

Y172
Isotype

IgG
Research areas
- Neuroscience
- Processes

Alternative Versions
- Anti-c-Jun (phospho S63) antibody [Y172] - BSA and Azide free (ab227533)
Assay kits
- c-Jun Transcription Factor Assay Kit (Colorimetric) (ab207195)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- NIH/3T3 whole cell lysate (ab7179)
Recombinant Protein
- Recombinant Human c-Jun protein (ab84134)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32385 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB	(2)	1/1000 - 1/10000. Detects a band of approximately 42 kDa (predicted molecular weight: 36 kDa).
IHC-P		1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/50 - 1/100 The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.
ICC/IF		1/100 - 1/200.
Dot blot		1/1000.
ELISA		Use at an assay dependent concentration.

Notes
WB 1/1000 - 1/10000. Detects a band of approximately 42 kDa (predicted molecular weight: 36 kDa).
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/50 - 1/100 The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.
ICC/IF 1/100 - 1/200.
Dot blot 1/1000.
ELISA Use at an assay dependent concentration.

Application notes

Is unsuitable for Flow Cyt.

Function

Transcription factor that recognizes and binds to the enhancer heptamer motif 5'-TGA[CG]TCA-3'. Promotes activity of NR5A1 when phosphorylated by HIPK3 leading to increased steroidogenic gene expression upon cAMP signaling pathway stimulation. Involved in activated KRAS-mediated transcriptional activation of USP28 in colorectal cancer (CRC) cells (PubMed:24623306). Binds to the USP28 promoter in colorectal cancer (CRC) cells (PubMed:24623306).
Sequence similarities

Belongs to the bZIP family. Jun subfamily.
Contains 1 bZIP (basic-leucine zipper) domain.
Post-translational
modifications

Ubiquitinated by the SCF(FBXW7), leading to its degradation. Ubiquitination takes place following phosphorylation, that promotes interaction with FBXW7.
Phosphorylated by CaMK4 and PRKDC; phosphorylation enhances the transcriptional activity. Phosphorylated by HIPK3. Phosphorylated by DYRK2 at Ser-243; this primes the protein for subsequent phosphorylation by GSK3B at Thr-239. Phosphorylated at Thr-239, Ser-243 and Ser-249 by GSK3B; phosphorylation reduces its ability to bind DNA. Phosphorylated by PAK2 at Thr-2, Thr-8, Thr-89, Thr-93 and Thr-286 thereby promoting JUN-mediated cell proliferation and transformation. Phosphorylated by PLK3 following hypoxia or UV irradiation, leading to increase DNA-binding activity.
Acetylated at Lys-271 by EP300.
Cellular localization

Nucleus.
Target information above from: UniProt accession P05412 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 280831 Cow
- Entrez Gene: 3725 Human
- Entrez Gene: 16476 Mouse
- Entrez Gene: 24516 Rat
- Omim: 165160 Human
- SwissProt: P05412 Human
- SwissProt: P05627 Mouse
- SwissProt: P17325 Rat
- Unigene: 696684 Human
- Unigene: 275071 Mouse
- Unigene: 93714 Rat
see all
Alternative names
- Activator protein 1 antibody
- AP 1 antibody
- AP-1 antibody
- AP1 antibody
- cJun antibody
- Enhancer Binding Protein AP1 antibody
- Jun Activation Domain Binding Protein antibody
- JUN antibody
- Jun oncogene antibody
- JUN protein antibody
- Jun proto oncogene antibody
- JUN_HUMAN antibody
- JUNC antibody
- Oncogene JUN antibody
- p39 antibody
- Proto oncogene c jun antibody
- Proto oncogene cJun antibody
- Proto-oncogene c-jun antibody
- Transcription Factor AP 1 antibody
- Transcription factor AP-1 antibody
- Transcription Factor AP1 antibody
- V jun avian sarcoma virus 17 oncogene homolog antibody
- V jun sarcoma virus 17 oncogene homolog (avian) antibody
- V jun sarcoma virus 17 oncogene homolog antibody
- V-jun avian sarcoma virus 17 oncogene homolog antibody
- vJun Avian Sarcoma Virus 17 Oncogene Homolog antibody
see all

Immunocytochemistry/ Immunofluorescence - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

Immunocytochemistry confocal image of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized anisomycin-treated NIH/3T3 cell line (mouse embryonic fibroblast), staining nuclear c-Jun with ab32385 at 1:500 dilution and ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1:1000 dilution. The counterstain was ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 dilution, and the nuclear counterstain was DAPI (blue).

The NIH/3T3 cells were treated with 250 ng/ml Anisomycin for 30 minutes and then the signal decreased after phosphatase treatment at 37°C for 2 hours.
Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

All lanes : Anti-c-Jun (phospho S63) antibody [Y172] (ab32385) at 0.1 µg/ml (purified)

Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 250 ng/ml anisomycin for 30 minutes whole cell lysates
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) treated with 250 ng/ml anisomycin for 30 minutes whole cell lysates. Then the membrane was incubated with phosphatase

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 36 kDa

Blocking and diluting buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast tissue sections labeling c-Jun with Purified ab32385 at 1:250 dilution (0.46 µg/ml). Heat mediated antigen retrieval was performed using using ab93684 (Tris/EDTA buffer, pH 9.0)ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylinwas used as a counterstain
Immunocytochemistry/ Immunofluorescence - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma cell line) cells labeling c-Jun (phospho S63) with ab32385 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing the expression was increased after treatment with anisomycin (1 µg/ml for 15 minutes), then decreased after treatment with the Lambda Protein Phosphatase treatment 31℃ for 2 hours. The nuclear counter stain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).
Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

All lanes : Anti-c-Jun (phospho S63) antibody [Y172] (ab32385) at 0.1 µg/ml (purified)

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1 ug/ml anisomycin for 15 minutes whole cell lysates
Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1 ug/ml anisomycin for 15 minutes whole cell lysates 15ug. Then the membrane was incubated with phosphatase.

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 36 kDa

Blocking and diluting buffer: 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32385).
Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

Lane 1 : Anti-c-Jun (phospho S63) antibody [Y172] (ab32385) at 1/1000 dilution (Unpurified)
Lanes 2-3 : Human HRPT2/Parafibromin peptide (ab23385) at 1/1000 dilution (Unpurified)

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates with NFDM/TBST
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1ug/mL anisomycin for 15 minutes whole cell lysates with NFDM/TBST
Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1ug/ml anisomycin for 15 minutes whole cell lysates. Then the membrane was incubated with phosphatase. with NFDM/TBST

Lysates/proteins at 15 µg per lane.

Blocking peptides at 5 % per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 36 kDa
ELISA - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

Antigen pS63:c-Jun (phospho S63); NP:c-Jun non-phospho. Antigen concentration 0.01~1 ng/ml.

Primary antibody concentration range 0~1000 ng/ml.

Secondary antibody is an Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG(H+L) used at a 1:2500 dilution.
Dot Blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

Unpurified ab32385 used at a 1:1000 dilution.
Secondary antibody is Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) used at a 1:100,000 dilution. Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
Lane 1: Human c-Jun (pS63) phospho peptide.
Lane 2: Human c-Jun non-phospho peptide.
Exposure time 3 minutes.
Western blot - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

All lanes : Anti-c-Jun (phospho S63) antibody [Y172] (ab32385) at 1/10000 dilution (Unpurified)

Lanes 1 & 3 : Untreated NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate
Lane 2 : NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate treated with ultraviolet light
Lane 4 : NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate treated with 25 µg/ml Anisomycin for 15 minutes at 37°C

Predicted band size: 36 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

Paraffin-embedded human breast carcinoma tissue stained for c-Jun (phospho S63) with unpurified ab32385 at a 1/50 dilution in immunohistochemical analysis.
Anti-c-Jun (phospho S63) antibody [Y172] (ab32385)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

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Datasheet download

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Publishing research using ab32385? Please let us know so that we can cite the reference in this datasheet.

ab32385 has been referenced in 68 publications.

Liang X & Ju J Matrine inhibits ovarian cancer cell viability and promotes apoptosis by regulating the ERK/JNK signaling pathway via p38MAPK. Oncol Rep 45:N/A (2021). PubMed: 33786627
Li L & Zhao W The mutual regulatory loop between TPTEP1 and miR-1303 in leukemogenesis of acute myeloid leukemia. Cancer Cell Int 21:260 (2021). PubMed: 33985519
Luo J et al. miR-576-5p promotes epithelial-to-mesenchymal transition in colorectal cancer by targeting the Wnt5a-mediated Wnt/ß-catenin signaling pathway. Mol Med Rep 23:N/A (2021). PubMed: 33300054
Jiao CY et al. BUB1B promotes extrahepatic cholangiocarcinoma progression via JNK/c-Jun pathways. Cell Death Dis 12:63 (2021). PubMed: 33431813
Xu X et al. YAP-TEAD up-regulates IRS2 expression to induce and deteriorate oesophageal cancer. J Cell Mol Med 25:2584-2595 (2021). PubMed: 33570213

View all Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

Application

Immunohistochemistry (Frozen sections)

Sample

Mouse Tissue sections (embryonic head mesenchyme)

Permeabilization

Yes - 0.1Triton in block buffer

Specification

embryonic head mesenchyme

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 13 2022

Application

Immunocytochemistry

Sample

Human Cultured Cells (SH-SY5Y cells)

Permeabilization

Yes - 0.1% Triton-X100 in PBS 10'

Specification

SH-SY5Y cells

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 20°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 08 2022

Application

Western blot

Sample

Mouse Cell lysate - whole cell (CT26 cells lysate)

Gel Running Conditions

Reduced Denaturing (4-12% Tris-Bis gel)

Loading amount

40 µg

Specification

CT26 cells lysate

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 06 2021

Application

Western blot

Sample

Chicken Cell lysate - whole cell (Chicken thrombocyte)

Gel Running Conditions

Reduced Denaturing (4-20% gradient)

Loading amount

10 µg

Specification

Chicken thrombocyte

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Mrs. Katie Elliott

Verified customer

Submitted Apr 25 2017

Question

no signal in WB with ab28807 species tested cow, mouse, human.

Abcam community

Verified customer

Asked on Jan 04 2012

Answer

Merci de nous avoir contactés. Nous sommes désolés d'apprendre que le produit que vous avez reçu ne fonctionne pas comme attendu. Je me permets de vous transférer le numéro de commande de remplacement gratuit de ce produit par une unité de ab32385 : *******. Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition. Nous serions ravis de recevoir vos résultats obtenus avec ab32385 chez le bovin sous forme d'une Abreview via le lien suivant : https://www.abcam.com/index.html?pageconfig=your_reviews&intAbID=32385&strAccessMode=public_create N'hésitez pas à me contacter de nouveau si vous avez d'autres questions.

Abcam Scientific Support

Answered on Jan 04 2012

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Assay kits

Compatible Secondaries

Isotype control

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (68)

Customer reviews and Q&As

Immunohistochemistry (Frozen sections) abreview for Anti-c-Jun (phospho S63) antibody [Y172]

Immunocytochemistry abreview for Anti-c-Jun (phospho S63) antibody [Y172]

Western blot abreview for Anti-c-Jun (phospho S63) antibody [Y172]

Western blot abreview for Anti-c-Jun (phospho S63) antibody [Y172]

Post-translational
modifications