Anti-Nuclear Matrix Protein p84 antibody [5E10]

• Flow Cyt

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Flow Cytometry - Anti-Nuclear Matrix Protein p84 antibody [5E10] (AB487)

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.

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• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Nuclear Matrix Protein p84 antibody [5E10] (AB487)

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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nuclear Matrix Protein p84 antibody [5E10] (AB487)

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nuclear Matrix Protein p84 antibody [5E10] (AB487)

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

• ICC/IF

AbReview15760****

Immunocytochemistry/ Immunofluorescence - Anti-Nuclear Matrix Protein p84 antibody [5E10] (AB487)

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.

This image is a courtesy of an anonymous Abreview

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Nuclear Matrix Protein p84 antibody [5E10] (AB487)

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.

• IP

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Immunoprecipitation - Anti-Nuclear Matrix Protein p84 antibody [5E10] (AB487)

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.

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All lanes:

Immunoprecipitation - Anti-Nuclear Matrix Protein p84 antibody [5E10] (ab487)

Predicted band size: 76 kDa

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• WB

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Western blot - Anti-Nuclear Matrix Protein p84 antibody [5E10] (AB487)

7.5% SDS-PAGE gel.

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All lanes:

Western blot - Anti-Nuclear Matrix Protein p84 antibody [5E10] (ab487) at 1/1000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 30 µg

Lane 2:

HeLa (human epithelial cell line from cervix adenocarcinoma) nuclear lysate at 30 µg

Secondary

All lanes:

HRP-conjugated anti-mouse IgG

Predicted band size: 76 kDa

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• WB

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Western blot - Anti-Nuclear Matrix Protein p84 antibody [5E10] (AB487)

All lanes:

Western blot - Anti-Nuclear Matrix Protein p84 antibody [5E10] (ab487) at 1/1000 dilution

All lanes:

Mouse cerebellum tissue lysates at 50 µg

Secondary

All lanes:

anti-mouse IgG HRP-conjugated antibody

Predicted band size: 47 kDa,50 kDa,68 kDa,70 kDa,71 kDa,73 kDa,74 kDa,76 kDa,77 kDa,78 kDa,80 kDa,82 kDa,88 kDa,90 kDa

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• WB

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Western blot - Anti-Nuclear Matrix Protein p84 antibody [5E10] (AB487)

All lanes:

Western blot - Anti-Nuclear Matrix Protein p84 antibody [5E10] (ab487) at 1/1000 dilution

Lane 1:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

HEK-293T whole cell lysate at 10 µg

Lane 3:

HEK-293T whole cell lysate at 5 µg

Secondary

All lanes:

anti-mouse IgG HRP-conjugated antibody

Predicted band size: 76 kDa

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