Anti-PRMT3 antibody [EPR13279]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRMT3 antibody [EPR13279] (AB191562)

Immunohistochemical analysis of paraffin-embedded, human cervix carcinoma tissue labeling PRMT3 with ab191562 at a 1/100 dilution. Counter stained with hematoxylin. In the negative control PBS was used instead of primary antibody.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PRMT3 antibody [EPR13279] (AB191562)

Immunofluorescence analysis of, paraformaldehyde-fixed, MCF7 cells labeling PRMT3 with ab191562 at a 1/50 dilution. As secondary antibody goat anti-rabbit IgG (Alexa Fluor^® 488) ab150077 was used at a 1/200 dilution. In blue DAPI staining. In the negative controls cells were treated with anti-PRMT3 at a 1/50 dilution as primary antibody and goat anti-mouse IgG (Alexa Fluor®594) at a 1/400 dilution as secondary antibody.

• IP

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Immunoprecipitation - Anti-PRMT3 antibody [EPR13279] (AB191562)

Western blot analysis on immunoprecipitation from 1) 293 cell lysate and 2) PBS, labeling PRMT3 using ab191562 at 1/40 dilution and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG at a 1/1500 dilution.

All lanes:

Immunoprecipitation - Anti-PRMT3 antibody [EPR13279] (ab191562)

Predicted band size: 60 kDa

Observed band size: 70 kDa

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• WB

Lab

Western blot - Anti-PRMT3 antibody [EPR13279] (AB191562)

Western blot : Anti-PRMT3 antibody [EPR13279] ab191562 staining at 1/2000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 68 kDa in Wild-type A549 cell lysates with no signal observed at this size in PRMT3 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PRMT3 antibody [EPR13279] (ab191562) at 1/2000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

PRMT3 knockout A549 at 20 µg

Lane 3:

HeLa at 20 µg

Lane 4:

U-87 MG at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 60 kDa

Observed band size: 68 kDa,37 kDa

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• WB

Supplier Data

Western blot - Anti-PRMT3 antibody [EPR13279] (AB191562)

All lanes:

Western blot - Anti-PRMT3 antibody [EPR13279] (ab191562) at 1/10000 dilution

Lane 1:

HepG2 cell lysate at 20 µg

Lane 2:

HeLa cell lysate at 20 µg

Lane 3:

293 cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Secondary

All lanes:

goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution

Predicted band size: 60 kDa

Observed band size: 70 kDa

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