MA1-013

antibody from Invitrogen Antibodies

Targeting: RAP1A KREV-1, SMGP21

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Immunohistochemistry (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-013

Provider

Invitrogen Antibodies

Product name

RAP1A Monoclonal Antibody (1D2-1C64)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

Western blot analysis of MA1-013 detects an ~21kDa protein.

Reactivity

Human, Mouse

Host

Mouse

Isotype

IgG

Antibody clone number

1D2-1C64

Vial size

100 µg

Concentration

1 mg/mL

Storage

-20°C

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Rap1 (green) showing staining in the in the cytoplasm of C2C12 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Rap1 monoclonal antibody (Product # MA1-013) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent 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 (Product # 37525) for 15 minutes at room temperature. Cells were probed with a Rap1 monoclonal antibody (Product # MA1-013) 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 (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation of Rap1 was performed on NIH3T3 cells. Antigen-antibody complexes were formed by incubating 750 µg of NIH3T3 cell lysate with 2 µg of a Rap1 monoclonal antibody (Product # MA1-013) overnight on a rocking platform at 4øC. The immune complexes were captured on 50 µL Protein A/G Plus Agarose (Product # 20423), washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). Eluted sample (right lane) and 25 µg 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 (Product # MA1-013) at a dilution of 1:500 overnight rotating at 4øC, washed in TBST, and probed with Clean-blot IP Detection Reagent (Product # 21230) at a dilution of 1:2500 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).