Anti-RPL9 antibody [EP13752]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-RPL9 antibody [EP13752] (AB182556)

HeLa cells were fixed with 100μl 4% PFA for 10 min at RT, then permeabilized with 100μl 90% methanol for 30 min (-20° C) followed by blocking with 10% goat serum for 1h at room temperature. Incubate ab182556 for 30 min at room temperature (RT) and then incubate Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) for 30 min at room temperature.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RPL9 antibody [EP13752] (AB182556)

HCT 116 cells were fixed in 4% PFA and permeabilized with 0.1% Triton X-100. Primary antibody, ab182556 at 1/100 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti- Rabbit secondary antibody (ab150081) at 1/1000 dilution at RT for 45 min. ab195889 AlexaFluor® 594-conjugated Anti-alpha Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with ab119348 overnight at 4° C. Nucleus were visualized using DAPI.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPL9 antibody [EP13752] (AB182556)

The human cerebrum sections were performed by Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins, then the sections were labelled with ab182556 at 1/1000 dilution for 30 mins at room temperature. The Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) were used as secondary antibody. The sections Counterstained with hematoxylin. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IP

Lab

Immunoprecipitation - Anti-RPL9 antibody [EP13752] (AB182556)

RPL9 was immunoprecipitated from 0.35 mg of HepG2 whole cell lysate with ab182556 at 1/30 dilution. Western blot was performed from the immunoprecipitated using ab182556 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPL9 antibody [EP13752] (AB182556)

The rat cerebrum sections were performed by Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins, then the sections were labelled with ab182556 at 1/5000 dilution for 30 mins at room temperature. The Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) were used as secondary antibody. The sections Counterstained with hematoxylin. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPL9 antibody [EP13752] (AB182556)

The mouse cerebrum sections were performed by Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins, then the sections were labelled with ab182556 at 1/5000 dilution for 30 mins at room temperature. The Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) were used as secondary antibody. The sections Counterstained with hematoxylin. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• WB

Lab

Western blot - Anti-RPL9 antibody [EP13752] (AB182556)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Primary antibody : incubated for 1h at room temperature. Secondary antibody : incubated for 1h at 1 : 20,000 at room temperature. Exposure time : Lane 1, Lane 3 : 3 seconds, Lane 2 : 10 seconds

All lanes:

Western blot - Anti-RPL9 antibody [EP13752] (ab182556) at 1/50000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 15 µg

Lane 3:

C6 (Rat glial tumor glial cell) whole cell lysate at 15 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 1/20000 dilution

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