|H M R Mk||Endogenous||34||Rabbit|
Western blot analysis of extracts from various cell lines using RPL5 Antibody.Learn more about how we get our images.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human RPL5 protein (hRPL5-Myc/DDK; +), using RPL5 Antibody.Learn more about how we get our images.
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised June 2016
Protocol Id: 263
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
RPL5 Antibody recognizes endogenous levels of total RPL5 protein.
Human, Mouse, Rat, Monkey
Bovine, Pig, Horse
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gln122 of human RPL5 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Ribosomal protein L5 (RPL5) is one of several proteins that comprise the 60S ribosomal subunit. RPL5 binds 5S rRNA and the nucleolar RPL11 protein to form the 5S ribonucleoprotein particle (RNP) that is incorporated into the large 60S ribosomal subunit (1). An RP-MDM2-p53 protein complex that contains ribosomal proteins RPL5, RPL11, and RPL23 acts as a nucleolar stress sensor that binds and inhibits MDM2 ubiquitin ligase activity and enhances p53-mediated transcriptional activity (2,3). RPL5 cooperates with RPL11 to influence ribosome biogenesis through regulating expression of the transcription factor c-Myc, which acts as the master regulator of ribosome biogenesis (4). Mutations in the corresponding RPL5 gene are associated with Diamond-Blackfan anemia, which is a form of red blood cell aplasia, and some cases of pediatric T-cell acute lymphoblastic leukemia (5,6).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|14568S||100 µl (10 western blots)||$ 255.0|