Overview

  • Product name
    Anti-RPS6 (phospho S235 + S236) antibody
    See all RPS6 primary antibodies
  • Description
    Rabbit polyclonal to RPS6 (phospho S235 + S236)
  • Specificity
    This antibody does not cross react with RPS6 phosphorylated on serines 244 and 247.
  • Tested applications
    Suitable for: WB, ICC/IF, Electron Microscopymore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide derived from the region of human RPS6 that contains serines 235 and 236 (unfortunately, the amino acid sequence is considered to be commercially sensitive)

  • Positive control
    • HeLa +/- TNF-alpha or anisomycin
  • General notes
    Centrifuge before opening to settle contents.

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: 0.05% Sodium Azide
    Constituents: Dulbecco's PBS without magnesium and calcium, 50% Glycerol, PBS, 1mg/ml BSA, pH 7.3
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated RPS6. The final product is generated by affinity chromatography using a RPS6-derived peptide that is phosphorylated at serines 235 and 236.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab12864 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
WB 1/1000. Detects a band of approximately 32 kDa (predicted molecular weight: 31 kDa).
ICC/IF Use at an assay dependent concentration. Used at a dilution of 1/100 for 2 hrs on HeLa cells (see Abreview submitted by Marco Biggiogera).
Electron Microscopy Use at an assay dependent dilution.

Target

Images



  • Predicted band size : 31 kDa
    Peptide Competition.
    Lysates prepared from HeLa cells left untreated (1) or treated with anisomycin (2-5) were resolved by SDS-PAGE on a 14% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer for one hour at room  temperature, and incubated with ab12864 for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphoserine-containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with a goat anti-rabbit IgG HRP conjugate secondary antibody and bands were detected using the Pierce SuperSignalTM method. The data show that only the peptide corresponding to RPS6 blocks the signal, verifying the specificity of the antibody. Peptide Competition. Lysates prepared from HeLa cells left untreated (1) or treated with anisomycin (2-5) were resolved by SDS-PAGE on a 14% po
  •  ab12864 at a 1/100 dilution staining human HeLa cells by ICC/IF. The cells were formaldehyde fixed and then incubated with the antibody for 2 hours. An Alexa Fluor ® 488 conjugated goat polyclonal antibody was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Fan G  et al. Danshensu alleviates cardiac ischaemia/reperfusion injury by inhibiting autophagy and apoptosis via activation of mTOR signalling. J Cell Mol Med 20:1908-19 (2016). Rat . Read more (PubMed: 27385290) »
  • Van Rechem C  et al. Lysine demethylase KDM4A associates with translation machinery and regulates protein synthesis. Cancer Discov 5:255-63 (2015). Human . Read more (PubMed: 25564516) »

See all 6 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse (macrophage, skeletal muscle) Human (kidney))
Gel Running Conditions
Reduced Denaturing (8-18% gradient)
Loading amount
5 µg
Treatment
293A cells were treated with mTORC1 catalytic inhibitor 50nM for 1hr to inhibit S6 phosphorylation
Specification
Mouse (macrophage, skeletal muscle) Human (kidney)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Jun 21 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Bos taurus Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing (4-15% Gradient)
Loading amount
20 µg
Specification
Liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Julie Kim

Verified customer

Submitted May 04 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (HeLa)
Loading amount
50 µg
Specification
HeLa
Blocking step
Milk as blocking agent for 15 minute(s) · Concentration: 5%
Username

Dr. Marco Biggiogera

Verified customer

Submitted Sep 15 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Formaldehyde
Username

Dr. Marco Biggiogera

Verified customer

Submitted Aug 09 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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