Antibodies, Proteins, Kits and Reagents for Life Science

328 339 ₸ Product size

100 µl 328 339 ₸

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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

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 & Species

Application	Species
Dot	Human
ICC/IF	Human
IHC-P	Human
WB	Mouse Human

See all applications and species data

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.
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

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 (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

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)

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