Overview

  • Product name

  • Description

    Rabbit polyclonal to RPS6
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Saccharomyces cerevisiae, Schizosaccharomyces pombe
    Predicted to work with: a wide range of other species
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human RPS6.

    Read Abcam's proprietary immunogen policy (Peptide available as ab42866.)

  • Positive control

    • This antibody gave a positive signal in the following whole cell lysates; HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) A431 (Human epithelial carcinoma cell line) HEK293 (Human embryonic kidney cell line) HepG2 (Human hepatocellular liver carcinoma cell line) MCF7 (Human breast adenocarcinoma cell line) SHSY-5Y (Human neuroblastoma cell line) IHC-P: FFPE human skin melanoma tissue sections. ICC/IF: A431 cell line

Properties

  • 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

    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab40820 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
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 31 kDa (predicted molecular weight: 28 kDa).
ICC/IF Use a concentration of 5 µg/ml.

Target

Images

  • All lanes : Anti-RPS6 antibody (ab40820) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate
    Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 7 : SHSY-5Y (Human neuroblastoma 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

    Performed under reducing conditions.

    Predicted band size: 28 kDa
    Observed band size: 31 kDa
    why is the actual band size different from the predicted?

  • All lanes : Anti-RPS6 antibody (ab40820) at 2 µg/ml

    Lane 1 : Whole cell lysate from mouse NIH-3T3 cell line
    Lane 2 : Whole cell lysate from mouse MEF cell line
    Lane 3 : Whole cell lysate from rat PC12 cell line

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated pig anti rabbit Ig at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 28 kDa


    Exposure time: 5 minutes


    Detection method: Supersignal West Dura

    See Abreview

  • IHC image of RPS6 staining in a section of formalin-fixed paraffin-embedded normal human skin melanoma* 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 20mins. The section was then incubated with ab40820, 5ug/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.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • ab40820 stained in A431 cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab40820 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

References

This product has been referenced in:

  • Yin X  et al. TFE3 fusions escape from controlling of mTOR signaling pathway and accumulate in the nucleus promoting genes expression in Xp11.2 translocation renal cell carcinomas. J Exp Clin Cancer Res 38:119 (2019). Read more (PubMed: 30849994) »
  • Liu Y  et al. Mitochondrial carrier protein overloading and misfolding induce aggresomes and proteostatic adaptations in the cytosol. Mol Biol Cell 30:1272-1284 (2019). Read more (PubMed: 30893019) »
See all 14 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (BY4741)
Gel Running Conditions
Reduced Denaturing (4-15%)
Loading amount
10 µg
Treatment
200ng/L Rapamycin for 1 hr
Specification
BY4741
Blocking step
Milk as blocking agent for 15 minute(s) · Concentration: 2% · Temperature: 25°C

Mai Sun

Verified customer

Submitted Dec 10 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Fruit fly (Drosophila melanogaster) Cell (S2 D.mel embryonic cell line)
Permeabilization
Yes - 0.05% Triton X100
Specification
S2 D.mel embryonic cell line
Fixative
Paraformaldehyde

Kelly Burke

Verified customer

Submitted Jul 22 2015

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Zebrafish Tissue lysate - whole (whole larvae)
Specification
whole larvae
Treatment
4% glucose, 4% fructose, 4% fructose + 100 nM Rapamycin
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 13 2013

Application
Western blot
Sample
Schizosaccharomyces pombe Cell lysate - whole cell (S.pombe whole cell lysate)
Loading amount
10 µg
Specification
S.pombe whole cell lysate
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Apr 05 2013

Application
Western blot
Sample
Leishmania Cell lysate - whole cell (L. amazonesis and L. donovani promastigotes)
Loading amount
50 µg
Specification
L. amazonesis and L. donovani promastigotes
Gel Running Conditions
Reduced Denaturing (15)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 11 2012

Answer

DISCOUNT CODE: 40497
Expiration date: 07/12/2012
I am very pleased to hear you would like to accept our offer and test ab40820 in Leishmania samples. This code will give you: 1 free PRIMARY ANTIBODY OR VALUE OFF OF ORDER before the expiration date. To redeem this offer, please submit an Abreview for Leishmania samples and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.
Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.
Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.
The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More
Application
Western blot
Sample
Mouse Cell lysate - whole cell (various incl: NIH-3T3, MEF, PC12)
Loading amount
30 µg
Specification
various incl: NIH-3T3, MEF, PC12
Gel Running Conditions
Reduced Denaturing (12.5% SDS PAGE)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C

Mr. Antibody Solutions

Verified customer

Submitted Oct 07 2008

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