Phospho-c-Jun (S73)+JunD (S100) Recombinant Rabbit Monoclonal Antibody [JE65-46]
cat.: HA722475
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | JE65-46 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 40/45 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phosphopeptide corresponding to residues around Ser73 of human c-Jun. |
| Positive control: | HeLa cell lysate, HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate, NIH/3T3 cell lysate, NIH/3T3 treated with 25μg/mL anisomycin for 30 minutes cell lysate, RAW264.7 cell lysate, PC-12 cell lysate. |
| Subcellular location: | Nucleus. |
| Recommended Dilutions:
WB IF-Cell FC |
1:1,000 1:100 1:1,000 |
| Uniprot #: | SwissProt: P05412 Human | P17535 Human | P05627 Mouse | P15066 Mouse | P17325 Rat | P52909 Rat |
| Alternative names: | Activator protein 1 AP 1 AP-1 AP1 cJun Enhancer Binding Protein AP1 Jun Activation Domain Binding Protein JUN Jun oncogene JUN protein Jun proto oncogene JUN_HUMAN JUNC Oncogene JUN p39 Proto oncogene c jun Proto oncogene cJun Proto-oncogene c-jun Transcription Factor AP 1 Transcription factor AP-1 Transcription Factor AP1 V jun avian sarcoma virus 17 oncogene homolog V jun sarcoma virus 17 oncogene homolog (avian) V jun sarcoma virus 17 oncogene homolog V-jun avian sarcoma virus 17 oncogene homolog vJun Avian Sarcoma Virus 17 Oncogene Homolog Activator protein 1 AP 1 AP1 Jun D jun D proto oncogene Jund JunD FL isoform JUND_HUMAN Transcription factor jun D Transcription factor jun-D |
Images
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Fig1:
Western blot analysis of Phospho-c-Jun (S73)+JunD (S100) on different lysates with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (HA722475) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate Lane 3: HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 40/45 kDa Observed band size: 40/45 kDa Exposure time: 30 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722475) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of Phospho-c-Jun (S73)+JunD (S100) on different lysates with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (HA722475) at 1/1,000 dilution. Lane 1: NIH/3T3 cell lysate Lane 2: NIH/3T3 treated with 25μg/mL anisomycin for 30 minutes cell lysate Lane 3: RAW264.7 cell lysate Lane 4: PC-12 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 40/45 kDa Observed band size: 40/45 kDa Exposure time: 3 minutes; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722475) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig3:
Immunocytochemistry analysis of HeLa cells untreated / treated with 250ng/mL anisomycin for 30 minutes labeling Phospho-c-Jun (S73)+JunD (S100) with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (HA722475) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (HA722475) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Flow cytometric analysis of HeLa cells untreated (left) / treated with 250ng/mL anisomycin for 30 minutes (right) labeling Phospho-c-Jun (S73)+JunD (S100). Cells were fixed and permeabilized. Then stained with the primary antibody (HA722475, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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