![Western Blot: RAP1A Antibody (1D4-1A3) [NBP2-22527] - Analysis of 25 ug of various whole cell lysate per well.](http://images.novusbio.com/fullsize/RAP1A-Antibody-1D4-1A3-Western-Blot-NBP2-22527-img0006.jpg "Western Blot: RAP1A Antibody (1D4-1A3) [NBP2-22527] - Analysis of 25 ug of various whole cell lysate per well.")
| Reactivity | Hu, Mu, RtSpecies Glossary |
| Applications | WB, Func, ICC/IF, IHC, IP |
| Clone | 1D4-1A3 |
| Clonality | Monoclonal |
| Host | Mouse |
| Conjugate | Unconjugated |
| Concentration | 1 mg/ml |
| Immunogen | Recombinant full-length human Rap1 |
| Isotype | IgG |
| Clonality | Monoclonal |
| Host | Mouse |
| Gene | RAP1A |
| Purity | Protein A purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
| Dilutions |
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| Application Notes | Use in Proximity Ligation Assay reported in scientific literature (PMID:30002060). |
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| Publications |
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| Storage | Store at -20C. Avoid freeze-thaw cycles. |
| Buffer | PBS with 1 mg/ml BSA and 30% glycerol |
| Preservative | 0.05% Sodium Azide |
| Concentration | 1 mg/ml |
| Purity | Protein A purified |
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Secondary Antibodies |
Isotype Controls |
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![Immunocytochemistry/Immunofluorescence: RAP1A Antibody (1D4-1A3) [NBP2-22527] - Analysis of Rap1 (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% BSA/TBST for 15 minutes at room temperature. Cells were probed with a Rap1 monoclonal antibody at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye.](http://images.novusbio.com/fullsize/RAP1A-Antibody-1D4-1A3-Immunocytochemistry-Immunofluorescence-NBP2-22527-img0003.jpg "Immunocytochemistry/Immunofluorescence: RAP1A Antibody (1D4-1A3) [NBP2-22527] - Analysis of Rap1 (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% BSA/TBST for 15 minutes at room temperature. Cells were probed with a Rap1 monoclonal antibody at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye.")
![Immunohistochemistry-Paraffin: RAP1A Antibody (1D4-1A3) [NBP2-22527] - Analysis showing staining in the cytoplasm and membrane of human colon carcinoma (right) compared to a negative control without primary antibody (left).](http://images.novusbio.com/fullsize/RAP1A-Antibody-1D4-1A3-Immunohistochemistry-Paraffin-NBP2-22527-img0005.jpg "Immunohistochemistry-Paraffin: RAP1A Antibody (1D4-1A3) [NBP2-22527] - Analysis showing staining in the cytoplasm and membrane of human colon carcinoma (right) compared to a negative control without primary antibody (left).")
![Immunohistochemistry-Paraffin: RAP1A Antibody (1D4-1A3) [NBP2-22527] - Analysis showing staining in the cytoplasm and membrane of human kidney tissue (right) compared to a negative control without primary antibody (left).](http://images.novusbio.com/fullsize/RAP1A-Antibody-1D4-1A3-Immunohistochemistry-Paraffin-NBP2-22527-img0004.jpg "Immunohistochemistry-Paraffin: RAP1A Antibody (1D4-1A3) [NBP2-22527] - Analysis showing staining in the cytoplasm and membrane of human kidney tissue (right) compared to a negative control without primary antibody (left).")
![Immunoprecipitation: RAP1A Antibody (1D4-1A3) [NBP2-22527] - Analysis of Rap1 was performed on NIH3T3 cells. Antigen-antibody complexes were formed by incubating 750ug of NIH3T3 whole cell lysate with 2ug of a Rap1 monoclonal antibody overnight on a rocking platform at 4C. The immune complexes were captured on 50ul Protein A/G Plus Agarose, washed extensively, and eluted with 5X Lane Marker Reducing Sample BUffer. Eluted sample (right lane) and 25ug of NIH3T3 control lysate (left lane) were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween for at least 1 hour. The membrane was probed with a Rap1 monoclonal antibody at a dilution of 1:500 overnight rotating at 4C, washed in TBST, and probed with Clean-Blot IP Detection Reagent at a dilution of 1:2500 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura.](http://images.novusbio.com/fullsize/RAP1A-Antibody-1D4-1A3-Immunoprecipitation-NBP2-22527-img0002.jpg "Immunoprecipitation: RAP1A Antibody (1D4-1A3) [NBP2-22527] - Analysis of Rap1 was performed on NIH3T3 cells. Antigen-antibody complexes were formed by incubating 750ug of NIH3T3 whole cell lysate with 2ug of a Rap1 monoclonal antibody overnight on a rocking platform at 4C. The immune complexes were captured on 50ul Protein A/G Plus Agarose, washed extensively, and eluted with 5X Lane Marker Reducing Sample BUffer. Eluted sample (right lane) and 25ug of NIH3T3 control lysate (left lane) were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween for at least 1 hour. The membrane was probed with a Rap1 monoclonal antibody at a dilution of 1:500 overnight rotating at 4C, washed in TBST, and probed with Clean-Blot IP Detection Reagent at a dilution of 1:2500 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura.")
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