Recombinant Anti-c-Jun antibody [EP693Y] - BSA and Azide free (ab247284)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP693Y] to c-Jun - BSA and Azide free
- Suitable for: IHC-P, WB, IP, ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-c-Jun antibody [EP693Y] - BSA and Azide free
See all c-Jun primary antibodies -
Description
Rabbit monoclonal [EP693Y] to c-Jun - BSA and Azide free -
Host species
Rabbit -
Specificity
PBS only lot tested.
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Tested applications
Suitable for: IHC-P, WB, IP, ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK-293, MOJ/3T3 and PC-12 cell lysates;. IHC-P: Rat liver, mouse cerebrum and human cervix carcinoma tissues. ICC/IF: NIH/3T3 and HeLa cells. ICC KO: HEK293 cells (HEK293-JUN KO cells used as a negative cell line). Flow Cyt (intra): HEK-293 cells IP: NIH/3T3 cell lysate
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General notes
ab247284 is the carrier-free version of ab40766.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP693Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 647 Anti-c-Jun antibody [EP693Y] (ab210012)
- Alexa Fluor® 488 Anti-c-Jun antibody [EP693Y] (ab210013)
- PE Anti-c-Jun antibody [EP693Y] (ab303101)
- APC Anti-c-Jun antibody [EP693Y] (ab303102)
- HRP Anti-c-Jun antibody [EP693Y] (ab303103)
- Alexa Fluor® 594 Anti-c-Jun antibody [EP693Y] (ab310444)
- Alexa Fluor® 555 Anti-c-Jun antibody [EP693Y] (ab311972)
- Alexa Fluor® 568 Anti-c-Jun antibody [EP693Y] (ab312445)
- Anti-c-Jun antibody [EP693Y] (ab40766)
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Conjugation kits
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Isotype control
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab247284 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 |
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 39 kDa (predicted molecular weight: 39 kDa).
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Detects a band of approximately 39 kDa (predicted molecular weight: 39 kDa). |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
Target
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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
modificationsUbiquitinated 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. - Information by UniProt
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Database links
- 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
see all -
Alternative names
- Activator protein 1 antibody
- AP 1 antibody
- AP-1 antibody
see all
Images
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This data was developed using ab40766, the same antibody clone in a different buffer formulation.
ab40766 staining c-Jun in wild-type HEK293 cells (top panel) and c-Jun knockout HEK293 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab40766 at 1/250 dilution and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI. Image was taken with a high-content analysis system (Perkin Elmer, Operetta CLS™). -
This data was developed using ab40766, the same antibody clone in a different buffer formulation.
Purified ab40766 at 1/20 dilution (1µg) immunoprecipitating c-Jun in NIH/3T3 whole cell lysate.
Lane 1 (input): NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10µg
Lane 2 (+): ab40766 + NIH/3T3 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40766 in NIH/3T3 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 39 kDa -
All lanes : Anti-c-Jun antibody [EP693Y] (ab40766) at 1/1000 dilution (Purified)
Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate
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: 39 kDa
Observed band size: 39 kDaThis data was developed using ab40766, the same antibody clone in a different buffer formulation.
Blocking Buffer and concentration: 5% NFDM/TBST
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This data was developed using ab40766, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling c-Jun with Purified ab247284 at 1:50 dilution (4.06 ?g/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control. -
This data was developed using ab40766, the same antibody clone in a different buffer formulation.Intracellular Flow Cytometry analysis of HEK-293 (Human embryonic kidney epithelial cell) cells labeling c-Jun with Purified ab247284 at 1/20 dilution (10µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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This data was developed using ab40766, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling c-Jun with Purified ab247284 at 1:500 dilution (0.41 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
This data was developed using ab40766, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum tissue sections labeling c-Jun with Purified ab247284 at 1:500 dilution (0.41 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
This data was developed using ab40766, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue sections labeling c-Jun with Purified ab247284 at 1:500 dilution (0.41 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
All lanes : Anti-c-Jun antibody [EP693Y] (ab40766) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : Jun knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lysates/proteins at 40 µg per lane.
Predicted band size: 39 kDaThis data was developed using ab40766, the same antibody clone in a different buffer formulation.
Lanes 1 - 2: Merged signal (red and green). Green - ab40766 observed at 35 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab40766 was shown to specifically react with Jun in wild-type HEK-293 cells as signal was lost in Jun knockout cells. Wild-type and Jun knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab40766 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Protocols
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.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab247284 has not yet been referenced specifically in any publications.