Antibodies, Proteins, Kits and Reagents for Life Science

284 784 ₸ Product size

100 µl 284 784 ₸

Order now and get it on Tuesday March 09, 2021

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: WB, IHC-P, ICC/IF, Flow Cyt, IP, Dot blot
Reacts with: Mouse, Rat, Human

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 & Species

Application	Species
Flow Cyt	Mouse
ICC/IF	Mouse
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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: NIH/3T3 cells. IP: MCF7 grown in serum-free media overnight, then grown in 20% FBS media for 30 minutes, whole cell lysate.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20770
Isotype

IgG
Research areas

Western blot - Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] (ab215214)

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.
Immunocytochemistry/ Immunofluorescence - Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] (ab215214)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] (ab215214)

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.
Dot Blot - Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] (ab215214)

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.
Flow Cytometry - Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] (ab215214)

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.
Immunoprecipitation - Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] (ab215214)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] (ab215214)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] (ab215214)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] (ab215214)

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.
Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] (ab215214)

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