WB
Western blot analysis of JunD on different lysates with Rabbit anti-JunD antibody at 1/2,000 dilution. Lane 1: RAW264.7 cell lysate, Lane 2: C6 cell lysate, Lane 3: HeLa cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 30 seconds; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-JunD antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-JunD antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of C6 cells labeling JunD with Rabbit anti-JunD antibody 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-JunD antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of NIH/3T3 cells labeling JunD. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
JunD was immunoprecipitated from 0.2 mg HeLa cell lysate with Rabbit anti-JunD antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-JunD antibody at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input), Lane 2: Rabbit anti-JunD antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-JunD antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 59 secondsChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from BCL-2 cells with JunD antibody or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Product Name | JunD Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | The activator protein-1 (AP-1) transcription factor consists of either Jun/Jun homodimers or Fos/Jun heterodimeric complexes. Homo- and heterodimers bind to the TGACTCA consensus sequence present in numerous promoters and initially identified as the phorbol ester tumor promoter response element (TRE). Jun B and Jun D have been shown to be almost identical to c-Jun in their C-terminal regions, which are involved in dimerization and DNA binding, whereas their N-terminal domains, which are involved in transcriptional activation, diverge. All three form heterodimers among themselves and with c-Fos and other members of the Fos gene family. Studies suggest that the two forms of Jun D may be due to internal initiation of translation. |
| Immunogen | recombinant protein |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | ChIPFCICC/IFIHCIPWB |
| WB:1:2000-1:5000 IHC:1:1000 ICC/IF:1:100 FC:1:1000 IP:1-2μg/sample ChIP:Use 0.5~2 μg for 25 μg of chromatin. |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | Activator protein 1 antibody AP 1 antibody AP1 antibody Jun D antibody jun D proto oncogene antibody Jund antibody JunD FL isoform antibody JUND_HUMAN antibody Transcription factor jun D antibody Transcription factor jun-D antibody |
|---|---|
| Molecular Weight(MW) | 35kDa(Observed band size: 39/42kDa) |
| Cellular Localization | Nucleus. |
| SwissProt ID | P17535 |
|---|
WB
Western blot analysis of JunD on different lysates with Rabbit anti-JunD antibody at 1/2,000 dilution. Lane 1: RAW264.7 cell lysate, Lane 2: C6 cell lysate, Lane 3: HeLa cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 30 seconds; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-JunD antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-JunD antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of C6 cells labeling JunD with Rabbit anti-JunD antibody 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-JunD antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of NIH/3T3 cells labeling JunD. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
JunD was immunoprecipitated from 0.2 mg HeLa cell lysate with Rabbit anti-JunD antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-JunD antibody at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input), Lane 2: Rabbit anti-JunD antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-JunD antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 59 seconds
ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from BCL-2 cells with JunD antibody or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Application Notes | WB:1:2000-1:5000 IHC:1:1000 ICC/IF:1:100 FC:1:1000 IP:1-2μg/sample ChIP:Use 0.5~2 μg for 25 μg of chromatin. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
