Antibodies, Proteins, Kits and Reagents for Life Science

257 980 ₸ Product size

100 µl 257 980 ₸

Order now and get it on Tuesday March 09, 2021

Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR16586] to JunD (phospho S100) + c-Jun (phospho S73)
Suitable for: Dot blot, IP, IHC-P, WB
Reacts with: Mouse, Rat, Human

Product name

Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586]
See all JunD+c-Jun primary antibodies
Description

Rabbit monoclonal [EPR16586] to JunD (phospho S100) + c-Jun (phospho S73)
Host species

Rabbit

Tested Applications & Species

Application	Species
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa untreated and treated with 250 ng/ml Anisomycin for 30 minutes whole cell lysates; NIH/3T3 untreated and treated with 250 ng/ml Anisomycin for 30 minutes whole cell lysates; RAW 264.7 and PC-12 whole cell lysates. IHC-P: Human tonsil, mouse testis and rat liver tissues. IP: HeLa treated with 250ng/ml Anisomycin for 30 minutes whole cell extract.
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.

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

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR16586
Isotype

IgG
Research areas

Western blot - Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858)

All lanes : Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858) at 1/10000 dilution

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 250 ng/ml Anisomycin for 30 minutes whole cell lysate
Lane 2 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG,(H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 36 kDa
Observed band size: 40,45,48 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/dilution buffer: 5% NFDM/TBST.
Per the blast, ab178858 could recognize JunD (phospho Ser100) with 100% homology. Multi-bands are due to c-Jun (phospho Ser73) & JunD (phospho Ser100).
Dot Blot - Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858)

Dot blot analysis of JunD (phospho S100) + c-Jun (phospho S73) peptide (Lane 1) and non-phospho peptide (Lane 2) labeled using ab178858 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
Western blot - Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858)

Lanes 1 & 4-5 : Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
Lanes 2-3 : Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858) at 1/10000 dilution

Lanes 1 & 3 & 5 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lanes 2 & 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 250 ng/ml Anisomycin for 30 minutes whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG,(H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 36 kDa
Observed band size: 40,45 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/dilution buffer: 5% NFDM/TBST.
Western blot - Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858)

All lanes : Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858) at 1/10000 dilution

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 250 ng/ml Anisomycin for 30 minutes whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 250 ng/ml Anisomycin for 30 minutes, whole cell lysate treated with Alkaline Phosphatase

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG,(H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 36 kDa
Observed band size: 40,45 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/dilution buffer: 5% NFDM/TBST.
Western blot - Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858)

All lanes : Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858) at 1/10000 dilution

Lane 1 : NIH/3T3 (Mouse embryo fibroblast cell line) treated with 250 ng/ml Anisomycin for 30 minutes whole cell lysate
Lane 2 : Untreated NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG,(H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 36 kDa
Observed band size: 40,45 kDa why is the actual band size different from the predicted?

Exposure time: 1 minute

Blocking/dilution buffer: 5% NFDM/TBST.
Per the blast, ab178858 could recognize JunD (phospho Ser100) with 100% homology. Multi-bands are due to c-Jun (phospho Ser73) & JunD (phospho Ser100).
Western blot - Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858)

All lanes : Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858) at 1/1000 dilution

Lane 1 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG,(H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 36 kDa
Observed band size: 40,45 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/dilution buffer: 5% NFDM/TBST.
Per the blast, ab178858 could recognize JunD (phospho Ser100) with 100% homology. Multi-bands are due to c-Jun (phospho Ser73) & JunD (phospho Ser100).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling JunD (phospho S100) + c-Jun (phospho S73) with ab178858 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on lymphocytes and endothelial cells of Human tonsil is observed. Counter stained with Hematoxylin.

Negative Control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling JunD (phospho S100) + c-Jun (phospho S73) with ab178858 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on spermatogoniums and Leydig cells of mouse testis is observed. Counter stained with Hematoxylin.

Negative Control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling JunD (phospho S100) + c-Jun (phospho S73) with ab178858 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on bile duct epithelial cells while no staining on hepatocytes of rat liver is observed. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858)

JunD (phospho S100) + c-Jun (phospho S73) were immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma), treated with 250ng/ml Anisomycin for 30 minutes, whole cell extract with ab178858 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab178858 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.
Lane 1: HeLa, treated with 250ng/ml Anisomycin for 30 minutes, whole cell extract
Lane 2: PBS.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.

Per the blast, ab178858 could recognize JunD (phospho Ser100) with 100% homology. Multi-bands are due to c-Jun (phospho Ser73) & JunD (phospho Ser100)
Anti-JunD (phospho S100) + c-Jun (phospho S73) antibody [EPR16586] (ab178858)

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