Recombinant Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] (ab215214)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20770] to RPS6 (phospho S240 + S244)
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP, Dot blot
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RPS6 (phospho S240 + S244) antibody [EPR20770]
See all RPS6 primary antibodies -
Description
Rabbit monoclonal [EPR20770] to RPS6 (phospho S240 + S244) -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP, Dot blotmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MCF7, NIH/3T3 and PC-12 grown in serum-free media overnight, then grown in 20% FBS media for 30 minutes, whole cell lysates. IHC-P: Human breast cancer and placenta tissues; Mouse colon tissue; Rat kidney tissue. ICC/IF: NIH/3T3 cells. Flow Cyt (intra): NIH/3T3 cells. IP: MCF7 grown in serum-free media overnight, then grown in 20% FBS media for 30 minutes, whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20770 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab215214 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 |
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Flow Cyt (Intra) |
1/600.
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WB |
1/5000. Detects a band of approximately 32 kDa (predicted molecular weight: 29 kDa).
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IHC-P |
1/500 - 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
1/500 dilution for human and 1/2000 for mouse and rat. |
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ICC/IF |
1/100.
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IP |
1/30.
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Dot blot |
1/1000.
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Notes |
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Flow Cyt (Intra)
1/600. |
WB
1/5000. Detects a band of approximately 32 kDa (predicted molecular weight: 29 kDa). |
IHC-P
1/500 - 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. 1/500 dilution for human and 1/2000 for mouse and rat. |
ICC/IF
1/100. |
IP
1/30. |
Dot blot
1/1000. |
Target
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Function
May play an important role in controlling cell growth and proliferation through the selective translation of particular classes of mRNA. -
Sequence similarities
Belongs to the ribosomal protein S6e family. -
Post-translational
modificationsRibosomal protein S6 is the major substrate of protein kinases in eukaryote ribosomes. The phosphorylation is stimulated by growth factors, tumor promoting agents, and mitogens. It is dephosphorylated at growth arrest. - Information by UniProt
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Database links
- Entrez Gene: 6194 Human
- Entrez Gene: 20104 Mouse
- Entrez Gene: 667739 Mouse
- Entrez Gene: 29304 Rat
- Omim: 180460 Human
- SwissProt: P62753 Human
- SwissProt: P62754 Mouse
- SwissProt: P62755 Rat
see all -
Alternative names
- 40S ribosomal protein S6 antibody
- Air8 antibody
- NP33 antibody
see all
Images
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All lanes : Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] (ab215214) at 1/5000 dilution
Lane 1 : MCF7 (human breast adenocarcinoma cell line) grown in serum-free media overnight, whole cell lysate
Lane 2 : MCF7 grown in serum-free media overnight, then grown in 20% FBS media for 30 minutes, whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast cell line) grown in serum-free media for 4 hours, whole cell lysate
Lane 4 : NIH/3T3 grown in serum-free media for 4 hours, then grown in 20% FBS media for 30 minutes, whole cell lysate
Lane 5 : PC-12 (rat adrenal gland pheochromocytoma cell line) grown in serum-free media overnight, whole cell lysate
Lane 6 : PC-12 grown in serum-free media overnight, then grown in 20% FBS media for 30 minutes, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 29 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?Exposure times: Lanes 1,2: 1 second; Lanes 3-6: 5 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast cell line) cells labeling RPS6 (phospho S240 + S244) with ab215214 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasmic staining was increased after treatment with 20% FBS for 30 minutes on NIH/3T3 cells. Cells were FBS deprived for 4 hours before the treatment.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling RPS6 (phospho S240 + S244) with ab215214 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Cytoplasmic staining on human placenta (image A), no signal was detected when tissues were treated with alkaline phosphatase (image B). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Dot blot analysis of RPS6 (phospho S240 + S244) labeled with ab215214 at 1/1000 dilution.
Lane 1: RPS6 (phospho S240 + S244) peptide.
Lane 2: RPS6 (phospho S240) peptide.
Lane 3: RPS6 (phospho S244) peptide.
Lane 4: RPS6 non-phospho peptide.
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast cell line) grown in serum-free media for 4 hours, then grown in 20% FBS media for 30 minutes (red) or NIH/3T3 grown in serum-free media for 4 hours (green), cell line labeling RPS6 (phospho S240 + S244) with ab215214 at 1/600 dilution compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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RPS9 (phospho S240 + S244) was immunoprecipitated from 0.35 mg of MCF7 (human breast adenocarcinoma cell line), grown in serum-free media overnight, then grown in 20% FBS media for 30 minutes, lysate with ab215214 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab215214 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Lane 1: MCF7 grown in serum-free media overnight, then grown in 20% FBS media for 30 minutes, whole cell lysate 10 μg (Input).
Lane 2: ab215214 IP in MCF7 grown in serum-free media overnight, then grown in 20% FBS media for 30 minutes, whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab215214 in MCF7 grown in serum-free media overnight, then grown in 20% FBS media for 30 minutes, whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
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Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling RPS6 (phospho S240 + S244) with ab215214 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Cytoplasmic staining on human breast cancer (image A), no signal was detected when tissues were treated with alkaline phosphatase (image B). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling RPS6 (phospho S240 + S244) with ab215214 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Cytoplasmic staining on mouse colon (image A), no signal was detected when tissues were treated with alkaline phosphatase (image B). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling RPS6 (phospho S240 + S244) with ab215214 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Cytoplasmic staining on rat kidney (image A), no signal was detected when tissues were treated with alkaline phosphatase (image B). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (8)
ab215214 has been referenced in 8 publications.
- Sawaguchi S et al. Hypomyelinating Leukodystrophy 8 (HLD8)-Associated Mutation of POLR3B Leads to Defective Oligodendroglial Morphological Differentiation Whose Effect Is Reversed by Ibuprofen. Neurol Int 14:212-244 (2022). PubMed: 35225888
- Bai Y et al. Phosphorylation-mediated PI3K-Art signalling pathway as a therapeutic mechanism in the hydrogen-induced alleviation of brain injury in septic mice. J Cell Mol Med 26:5713-5727 (2022). PubMed: 36308410
- Hattori K et al. The Infantile Leukoencephalopathy-Associated Mutation of C11ORF73/HIKESHI Proteins Generates de novo Interactive Activity with Filamin A, Inhibiting Oligodendroglial Cell Morphological Differentiation. Medicines (Basel) 8:N/A (2021). PubMed: 33535532
- Sawaguchi S et al. Hypomyelinating Leukodystrophy 7 (HLD7)-Associated Mutation of POLR3A Is Related to Defective Oligodendroglial Cell Differentiation, Which Is Ameliorated by Ibuprofen. Neurol Int 14:11-33 (2021). PubMed: 35076634
- Xu M et al. Interaction of YAP1 and mTOR promotes bladder cancer progression. Int J Oncol 56:232-242 (2020). PubMed: 31789387
- Matsumoto N et al. PP1C and PP2A are p70S6K Phosphatases Whose Inhibition Ameliorates HLD12-Associated Inhibition of Oligodendroglial Cell Morphological Differentiation. Biomedicines 8:N/A (2020). PubMed: 32316234
- Zhou J et al. HDAC1-mediated deacetylation of LSD1 regulates vascular calcification by promoting autophagy in chronic renal failure. J Cell Mol Med 24:8636-8649 (2020). PubMed: 32596952
- Matsumoto N et al. Hypomyelinating leukodystrophy-associated mutation of RARS leads it to the lysosome, inhibiting oligodendroglial morphological differentiation. Biochem Biophys Rep 20:100705 (2019). PubMed: 31737794