Anti-RPL11 antibody

• WB

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Western blot - Anti-RPL11 antibody (AB79352)

RPL11 contains a number of potential phosphorylation and acetylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

All lanes:

Western blot - Anti-RPL11 antibody (ab79352) at 1 µg/mL

Lane 1:

Human liver tissue lysate - total protein (<a href='/en-us/products/unavailable/human-liver-tissue-lysate-total-protein-ab29889'>ab29889</a>) at 10 µg

Lane 2:

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

Lane 3:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 10 µg

Lane 4:

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

Lane 5:

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

Lane 6:

Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 7:

HEK293 (Human embryonic kidney 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: 20 kDa

Observed band size: 22 kDa,30 kDa,45 kDa,70 kDa

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPL11 antibody (AB79352)

IHC image of ab79352 staining in human pancreas formalin fixed paraffin embedded tissue section, 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 20 mins. The section was then incubated with ab79352, 1µg/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.

• IP

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Immunoprecipitation - Anti-RPL11 antibody (AB79352)

RPL11 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to RPL11 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab79352.
Secondary : Clean blot (HRP conjugate) at 1/1000 dilution.
Band : 22kDa : RPL11

All lanes:

Immunoprecipitation - Anti-RPL11 antibody (ab79352)

Predicted band size: 20 kDa

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• WB

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Western blot - Anti-RPL11 antibody (AB79352)

All lanes:

Western blot - Anti-RPL11 antibody (ab79352) at 1 µg/mL

Lane 1:

Marker

Lane 2:

Zebrafish brain homogenate at 20 µg

Lane 3:

Zebrafish heart homogenate at 20 µg

Lane 4:

Zebrafish liver homogenate at 20 µg

Lane 5:

Zebrafish skeletal muscle homogenate at 20 µg

Lane 6:

Human hepatocellular carcinoma cell line at 20 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution

Predicted band size: 20 kDa

Observed band size: 22 kDa

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Exposure time: 3min

• WB

CiteAb

Western blot - Anti-RPL11 antibody (AB79352)

RPL11 western blot using anti-RPL11 antibody ab79352. Publication image and figure legend from Hao, Q., Wang, J., et al., 2020, Cell Death Dis, PubMed 32198344.

ab79352 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab79352 please see the product overview.

Ablation of endogenous SBDS induces RPL5- and RPL11-dependent p53 stabilization.a, b Knockdown of SBDS prolongs p53 protein half-life. HCT116p53+/+ cells were transfected with control or SBDS siRNA for 48–72 hr. CHX was supplemented into the medium for the indicated time before the cells were harvested for IB analysis a. Quantification of the p53/β-actin ratios is shown in the b. c RPL5 and RPL11 are required for SBDS depletion-induced p53 activation. HCT116p53+/+ cells were transfected with the indicated siRNAs for 48–72 hr and harvested for IB analysis using the antibodies as indicated. d Knockdown of SBDS enhances the RPL11-MDM2 interaction. HCT116p53+/+ cells were transfected with control or SBDS siRNA for ~48 hr. The proteasome inhibitor MG132 was supplemented into the medium for 4 hr before the cells were harvested for co-IP-IB assays using antibodies as indicated. e Knockdown of SBDS enhances the RPL5-MDM2 interaction. The same experiments were conducted as described in d, except that the anti-RPL5 antibody was used.

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