Anti-RNF20 antibody [RM1054] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RNF20 antibody [RM1054] - BSA and Azide free (AB314770)

This data was developed using ab314769, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling RNF20 with ab314769 at 1/500 (0.902 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing nuclear staining in HeLa cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-RNF20 antibody [RM1054] - BSA and Azide free (AB314770)

This data was developed using ab314769, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling RNF20 with ab314769 at 1/50 dilution (1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• WB

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Western blot - Anti-RNF20 antibody [RM1054] - BSA and Azide free (AB314770)

This data was developed using ab314769, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. The identity of the lower MW band at approximately 37 kDa (in lane 3) is unknown. Lanes 1 and lane 2 blot were developed using a high sensitivity ECL substrate. Samples are non-boiled as boiling may cause protein aggregation. Exposure time : Lanes 1 and 2 : 180 seconds; Lane 3 : 103 seconds

All lanes:

Western blot - Anti-RNF20 antibody [RM1054] (<a href='/en-us/products/primary-antibodies/rnf20-antibody-rm1054-ab314769'>ab314769</a>) at 1/1000 dilution

Lane 1:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 114 kDa

true

• WB

Supplier Data

Western blot - Anti-RNF20 antibody [RM1054] - BSA and Azide free (AB314770)

This data was developed using ab314769, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. The identity of the lower MW band at approximately 37 kDa is unknown. In western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-RNF20 antibody [RM1054] (<a href='/en-us/products/primary-antibodies/rnf20-antibody-rm1054-ab314769'>ab314769</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting RNF20 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 114 kDa

false

Exposure time: 180s