Anti-RPL5 antibody (ab86863)
Key features and details
- Rabbit polyclonal to RPL5
- Suitable for: IP, IHC-P, WB, ICC/IF
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-RPL5 antibody
See all RPL5 primary antibodies -
Description
Rabbit polyclonal to RPL5 -
Host species
Rabbit -
Tested applications
Suitable for: IP, IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide corresponding to Human RPL5 aa 200 to the C-terminus conjugated to keyhole limpet haemocyanin.
(Peptide available asab97494) -
Positive control
- This antibody gave a positive signal Human fetal liver tissue lysate as well as the following whole cell lysates: HepG2; MEL-1; Jurkat.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab86863 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application Abreviews Notes IP Use a concentration of 5 µg/ml. IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. WB Use a concentration of 1 µg/ml. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa). ICC/IF Use a concentration of 5 µg/ml. Target
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Function
Required for rRNA maturation and formation of the 60S ribosomal subunits. This protein binds 5S RNA. -
Involvement in disease
Defects in RPL5 are the cause of Diamond-Blackfan anemia type 6 (DBA6) [MIM:612561]. DBA6 is a form of Diamond-Blackfan anemia, a congenital non-regenerative hypoplastic anemia that usually presents early in infancy. Diamond-Blackfan anemia is characterized by a moderate to severe macrocytic anemia, erythroblastopenia, and an increased risk of malignancy. 30 to 40% of Diamond-Blackfan anemia patients present with short stature and congenital anomalies, the most frequent being craniofacial (Pierre-Robin syndrome and cleft palate), thumb and urogenital anomalies. -
Sequence similarities
Belongs to the ribosomal protein L18P family. -
Cellular localization
Cytoplasm. Nucleus > nucleolus. - Information by UniProt
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Database links
- Entrez Gene: 6125 Human
- Omim: 603634 Human
- SwissProt: P46777 Human
- Unigene: 532359 Human
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Alternative names
- 60S ribosomal protein L5 antibody
- DBA6 antibody
- L5 antibody
see all
Images
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Western blot - Anti-RPL5 antibody (ab86863)All lanes : Anti-RPL5 antibody (ab86863) at 1 µg/ml
Lane 1 : Liver (Human)Tissue Lysate - Fetal tissue
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : MEL-1 (Human embryonic stem cell, male cell line) Whole Cell Lysate (ab27198)
Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 34 kDa
Additional bands at: 68 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute -
Immunocytochemistry/ Immunofluorescence - Anti-RPL5 antibody (ab86863)ICC/IF image of ab86863 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab86863, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, HepG2 and MCF7 cells at 5µg/ml -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPL5 antibody (ab86863)IHC image of RPL5 staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab86863, 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. -
Immunoprecipitation - Anti-RPL5 antibody (ab86863)RPL5 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to RPL5 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, HepG2 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab86863.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 34kDa, non specific band - 68kDa as present in test (lane 1) and control (lane 2); 68kDa: We are unsure as to the identity of this extra band; RPL5
Protocols
Datasheets and documents
References (6)
ab86863 has been referenced in 6 publications.
- Montellese C et al. USP16 counteracts mono-ubiquitination of RPS27a and promotes maturation of the 40S ribosomal subunit. Elife 9:N/A (2020). PubMed: 32129764
- Watanabe S et al. GRWD1 regulates ribosomal protein L23 levels via the ubiquitin-proteasome system. J Cell Sci 131:N/A (2018). PubMed: 29991511
- O'Brien R et al. MYC-Driven Neuroblastomas Are Addicted to a Telomerase-Independent Function of Dyskerin. Cancer Res 76:3604-17 (2016). PubMed: 27197171
- Wandrey F et al. The NF45/NF90 Heterodimer Contributes to the Biogenesis of 60S Ribosomal Subunits and Influences Nucleolar Morphology. Mol Cell Biol 35:3491-503 (2015). PubMed: 26240280
- Goudarzi KM et al. mTOR inhibitors blunt the p53 response to nucleolar stress by regulating RPL11 and MDM2 levels. Cancer Biol Ther 15:1499-514 (2014). Human . PubMed: 25482947
- Wu H et al. Human RNase H1 is associated with protein P32 and is involved in mitochondrial pre-rRNA processing. PLoS One 8:e71006 (2013). IP . PubMed: 23990920
Images
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Western blot - Anti-RPL5 antibody (ab86863)All lanes : Anti-RPL5 antibody (ab86863) at 1 µg/ml
Lane 1 : Liver (Human)Tissue Lysate - Fetal tissue
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : MEL-1 (Human embryonic stem cell, male cell line) Whole Cell Lysate (ab27198)
Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 34 kDa
Additional bands at: 68 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
-
Immunocytochemistry/ Immunofluorescence - Anti-RPL5 antibody (ab86863)ICC/IF image of ab86863 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab86863, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, HepG2 and MCF7 cells at 5µg/ml
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPL5 antibody (ab86863)IHC image of RPL5 staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab86863, 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.
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Immunoprecipitation - Anti-RPL5 antibody (ab86863)RPL5 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to RPL5 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, HepG2 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab86863.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 34kDa, non specific band - 68kDa as present in test (lane 1) and control (lane 2); 68kDa: We are unsure as to the identity of this extra band; RPL5
