RAP1A Antibody (1D2-1C64)

Western Blot: RAP1A Antibody (1D2-1C64) [NBP2-22526] - Analysis of 25 ug of various whole cell lysates per well.

Immunocytochemistry/Immunofluorescence: RAP1A Antibody (1D2-1C64) [NBP2-22526] - Analysis of Rap1 (green) showing staining in the in the cytoplasm of C2C12 cells (right) compared to a negative control without primary antibody (left).

Immunohistochemistry: RAP1A Antibody (1D2-1C64) [NBP2-22526] - Analysis of Rap1 showing staining in the membrane of human kidney tissue (right) compared with a negative control without primary antibody (left).

Immunocytochemistry/Immunofluorescence: RAP1A Antibody (1D2-1C64) [NBP2-22526] - 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.

Immunoprecipitation: RAP1A Antibody (1D2-1C64) [NBP2-22526] - Analysis of Rap1 was performed on NIH3T3 cells. Antigen-antibody complexes were formed by incubating 750ug of NIH3T3 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.

• Reactivity: Hu, Mu
• Applications: WB, ICC/IF, IHC, IP
• Clone: 1D2-1C64
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

RAP1A Antibody (1D2-1C64) Summary

• Immunogen: Recombinant full-length human Rap1
• Specificity: This product was made specifically to the RAP1A human protein (P62834). It has not yet been tested for cross reactivity to other related proteins.
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: RAP1A
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:10 - 1:200
  • Immunohistochemistry 1:100 - 1:1000
  • Immunohistochemistry-Paraffin 1:100 - 1:1000
  • Immunoprecipitation 2 ug
  • Western Blot 1:500 - 1:1000
• Theoretical MW: 21 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

Packaging, Storage & Formulations

• 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

Alternate Names for RAP1A Antibody (1D2-1C64)

• C21KG
• G-22K
• GTP-binding protein smg p21A
• KREV-1
• KREV1Ras-related protein Krev-1
• RAP1
• Rap1A
• RAP1A, member of RAS oncogene family
• RAS-related protein RAP1A
• ras-related protein Rap-1A
• SMGP21

Background

The product of this gene belongs to the family of RAS-related proteins. These proteins share approximately 50% amino acid identity with the classical RAS proteins and have numerous structural features in common. The most striking difference between RAP proteins and RAS proteins resides in their 61st amino acid: glutamine in RAS is replaced by threonine in RAP proteins. The product of this gene counteracts the mitogenic function of RAS because it can interact with RAS GAPs and RAF in a competitive manner. Two transcript variants encoding the same protein have been identified for this gene.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: RAP1A
• Uniprot:
  • P62834()