Anti-Nuclear Matrix蛋白p84抗体[5E10] (ab487)

Immunohistochemical analysis of human lung cancer tissue labeling Nuclear Matrix Protein p84 at the nucleus with ab487 at 1/100 dilution. 

Immunocytochemical analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Nuclear Matrix Protein p84 at the nucleus with ab487 at 1/500 dilution. Red: phalloidin, a cytoskeleton marker, diluted at 1:100.

Nuclear Matrix Protein p84 was immunoprecipitated from HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate with 3 µg ab487. Western blot was performed from the immunoprecipitate using ab487. Anti-Rabbit IgG was used as a secondary reagent.

Lane 1: HepG2 whole cell lysate 30 μg.

Lane 2: Control IP in HepG2 whole cell lysate with 3 μg of pre-immune mouse IgG.

Lane 3: ab487 IP in HepG2 whole cell lysate.

This image was generated using the ascites version of the product.

Overlay histogram showing HeLa cells stained with ab487 (red line). The cells were fixed with 100% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab487, 1µg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

This image was generated using the ascites version of the product.

7.5% SDS-PAGE gel.

This image was generated using the ascites version of the product.

Immunohistochemical analysis of human breast carcinoma tissue labeling Nuclear Matrix Protein p84 at the nucleus with ab487 at 1/200 dilution. 

ab487 staining Nuclear Matrix Protein p84 in Human stomach adenocarcinoma cell line (AGS) by Immunocytochemistry/ Immunofluorescence. The cells were formaldehyde fixed, permeabilised in 0.025% Triton X-100, TBS and then blocked using 5% serum for 1 hour at 23°C. Samples were then incubated with primary antibody at 2µg/ml for 1 hour at 23°C. The secondary antibody used was a goat anti-mouse IgG conjugated to Alexa Fluor® 350 (blue) used undiluted.p84 shows nuclear localization.

This image was generated using the ascites version of the product.

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ICC/IF image of ab487 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab487, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

This image was generated using the ascites version of the product.

7.5% SDS-PAGE gel.

This image was generated using the ascites version of the product.