Anti-RPS6 antibody - Ribosome Marker

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPS6 antibody - Ribosome Marker (AB40820)

IHC image of RPS6 staining in a section of formalin-fixed paraffin-embedded normal human skin melanoma* performed on a Leica BOND^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab40820, 5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RPS6 antibody - Ribosome Marker (AB40820)

ab40820 stained in A431 cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab40820 at 5μg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 μM for 1hour at room temperature.

• WB

Project

Western blot - Anti-RPS6 antibody - Ribosome Marker (AB40820)

All lanes:

Western blot - Anti-RPS6 antibody - Ribosome Marker (ab40820) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Lane 3:

A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

HEK293 Human embryonic kidney cell line Whole Cell Lysate at 10 µg

Lane 5:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 6:

MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Lane 7:

SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 29 kDa

Observed band size: 31 kDa

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• WB

AbReview12264****

Western blot - Anti-RPS6 antibody - Ribosome Marker (AB40820)

Detection method : Supersignal West Dura

All lanes:

Western blot - Anti-RPS6 antibody - Ribosome Marker (ab40820) at 2 µg/mL

Lane 1:

Whole cell lysate from mouse NIH-3T3 cell line at 30 µg

Lane 2:

Whole cell lysate from mouse MEF cell line at 30 µg

Lane 3:

Whole cell lysate from rat PC12 cell line at 30 µg

Secondary

All lanes:

HRP-conjugated pig anti rabbit Ig at 1/5000 dilution

Predicted band size: 29 kDa

false

Exposure time: 5min

This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.