Anti-RAP1A antibody [1D2-1C64]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RAP1A antibody [1D2-1C64] (AB175329)

Immunofluorescence analysis of formalin-fixed permeabilized HeLa cells, labeling RAP1A (green, left panel) using ab175329 at a 1/100 dilution followed by DyLight 488-conjugated goat anti-mouse IgG secondary antibody at a 1/400 dilution. Nuclei (blue) were stained with Hoechst 33342 dye (central panel).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAP1A antibody [1D2-1C64] (AB175329)

Immunohistochemistry analysis of RAP1A using ab175329 at 1/200 dilution showing staining in the membrane of paraffin-embedded human kidney tissue (right) compared with a negative control without primary antibody (left). Detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit.

Antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RAP1A antibody [1D2-1C64] (AB175329)

Immunocytochemical analysis of RAP1A (green) showing staining in the in the cytoplasm of Formalin-fixed and 0.1% Triton X-100 permeabilized C2C12 cells (right) using ab175329 at 1/20 dilution compared to a negative control without primary antibody (left) followed by DyLight-conjugated secondary antibody. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with DAPI.

• IP

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Immunoprecipitation - Anti-RAP1A antibody [1D2-1C64] (AB175329)

Western blot analysis on immunoprecipitation pellet from mouse NIH 3T3 cells. The antigen-antibody complex was formed by incubating 750 μg of NIH 3T3 whole cell lysate with 2 μg of ab175329 overnight at 4°C. The immune-complex was then captured on 50 μl Protein A/G Plus Agarose, washes extensively and eluted in sample buffer. 1) 25 μg of NIH 3T3 whole cell lysate, as a control, and 2) eluted sample were resolved on a SDS PAGE gel. The membrane was probed with ab175329 at a 1/500 dilution. Chemiluminescent detection was perfomed.

All lanes:

Immunoprecipitation - Anti-RAP1A antibody [1D2-1C64] (ab175329)

Predicted band size: 21 kDa

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• WB

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Western blot - Anti-RAP1A antibody [1D2-1C64] (AB175329)

All lanes:

Western blot - Anti-RAP1A antibody [1D2-1C64] (ab175329) at 1/500 dilution

Lane 1:

HeLa cell lysate at 25 µg

Lane 2:

HEK293T cell lysate at 25 µg

Lane 3:

U2OS cell lysate at 25 µg

Lane 4:

Mouse NIH 3T3 cell lysate at 25 µg

Secondary

All lanes:

Goat anti-mouse IgG-HRP at 1/15000 dilution

Predicted band size: 21 kDa

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