Overview

  • Product name
    Anti-SAM68 antibody
    See all SAM68 primary antibodies
  • Description
    Rabbit polyclonal to SAM68
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 400 to the C-terminus of Human SAM68.

    Read Abcam's proprietary immunogen policy

    (Peptide available as ab27829.)

  • Positive control
    • ab26803 gave a positive result in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) NIH 3T3 (Mouse embryonic fibroblast cell line) MEF1 (Mouse embryonic fibroblast cell line) PC12 (Rat adrenal pheochromocytoma cell line)

Properties

Applications

Our Abpromise guarantee covers the use of ab26803 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
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 68 kDa (predicted molecular weight: 49 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function
    Recruited and tyrosine phosphorylated by several receptor systems, for example the T-cell, leptin and insulin receptors. Once phosphorylated, functions as an adapter protein in signal transduction cascades by binding to SH2 and SH3 domain-containing proteins. Role in G2-M progression in the cell cycle. Represses CBP-dependent transcriptional activation apparently by competing with other nuclear factors for binding to CBP. Also acts as a putative regulator of mRNA stability and/or translation rates and mediates mRNA nuclear export.
    Isoform 3, which is expressed in growth-arrested cells only, inhibits S phase.
  • Tissue specificity
    Ubiquitously expressed in all tissue examined. Isoform 1 is expressed at lower levels in brain, skeletal muscle, and liver whereas isoform 3 is intensified in skeletal muscle and in liver.
  • Sequence similarities
    Belongs to the KHDRBS family.
    Contains 1 KH domain.
  • Developmental stage
    Isoform 3 is only expressed in growth-arrested cells.
  • Domain
    The KH domain is required for binding to RNA.
    The Pro-rich domains are flanked by Arg/Gly-rich motifs which can be asymmetric dimethylated on arginine residues to give the DMA/Gly-rich regions. Selective methylation on these motifs can modulate protein-protein interactions.
  • Post-translational
    modifications
    Tyrosine phosphorylated by several non-receptor tyrosine kinases, for example LCK, FYN and JAK3. Negatively correlates with ability to bind RNA but required for many interactions with proteins.
    Acetylated. Positively correlates with ability to bind RNA.
    Arginine methylation is required for nuclear localization. Also can affect interaction with other proteins. Inhibits interaction with Src-like SH3 domains, but not interaction with WW domains of WBP4/FBP21 AND FNBP4/FBP30.
    Arg-291, Arg-331 and Arg-346 are found to be also dimethylated, probably to asymmetric dimethylarginine.
  • Cellular localization
    Nucleus. Membrane.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • FLJ34027 antibody
    • GAP associated tyrosine phosphoprotein p62 antibody
    • GAP-associated tyrosine phosphoprotein p62 antibody
    • KH domain containing RNA binding signal transduction associated 1 antibody
    • KH domain-containing antibody
    • KHDR1_HUMAN antibody
    • Khdrbs1 antibody
    • p21 Ras GTPase activating protein associated p62 antibody
    • p21 Ras GTPase-activating protein-associated p62 antibody
    • p62 antibody
    • p68 antibody
    • RNA-binding antibody
    • Sam68 antibody
    • signal transduction-associated protein 1 antibody
    • Src associated in mitosis 68 kDa protein antibody
    • Src-associated in mitosis 68 kDa protein antibody
    see all

Images

  • Western blot - Anti-SAM68 antibody (ab26803)
    Western blot - Anti-SAM68 antibody (ab26803)
    All lanes : Anti-SAM68 antibody (ab26803) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 49 kDa
    Observed band size: 68 kDa (why is the actual band size different from the predicted?)



    Although the calculated molecular weight of SAM68 is 49 kDa, it has been shown to migrate at 68 kDa (see Molecular Biology of the Cell 14(1):274). ab26803 detects a band that is 68 kDa.
  • Western blot - Anti-SAM68 antibody (ab26803)
    Western blot - Anti-SAM68 antibody (ab26803)
    All lanes : Anti-SAM68 antibody (ab26803) at 1 µg/ml

    Lane 1 : NIH 3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Predicted band size: 49 kDa
    Observed band size: 68 kDa (why is the actual band size different from the predicted?)



    Although the calculated molecular weight of SAM68 is 49 kDa, it has been shown to migrate at 68 kDa (see Molecular Biology of the Cell 14(1):274). ab26803 detects a band that is 68 kDa.

  • Immunocytochemistry/ Immunofluorescence - Anti-SAM68 antibody (ab26803)
    Immunocytochemistry/ Immunofluorescence - Anti-SAM68 antibody (ab26803)

    ICC/IF image of ab26803 stained human HEK 293 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab26803, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, HepG2 and MCF7 cells.

  • Western blot - Anti-SAM68 antibody (ab26803)
    Western blot - Anti-SAM68 antibody (ab26803)This image is courtesy of an anonymous Abreview
    Anti-SAM68 antibody (ab26803) at 1/600 dilution (for 20 hours at 4°C in blocking buffer) + Whole cell lysate of human fibroblasts at 20 µg

    Secondary
    An IRDye®800CW Donkey anti-rabbit IgG polyclonal at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 49 kDa
    Observed band size: 68 kDa (why is the actual band size different from the predicted?)


    Exposure time: 1 minute


    Blocking Step: 5% BSA for 1 hour at 25°C

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SAM68 antibody (ab26803)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SAM68 antibody (ab26803)
    IHC image of ab26803 staining in human skin 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 ab26803, 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.

    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.

References

This product has been referenced in:
  • Puckette M  et al. Foot-and-Mouth Disease (FMD) Virus 3C Protease Mutant L127P: Implications for FMD Vaccine Development. J Virol 91:N/A (2017). Read more (PubMed: 28878081) »
  • Palmer SJ  et al. GTF2IRD2 from the Williams-Beuren critical region encodes a mobile-element-derived fusion protein that antagonizes the action of its related family members. J Cell Sci 125:5040-50 (2012). ICC/IF ; Mouse, Human . Read more (PubMed: 22899722) »

See all 2 Publications for this product

Publishing research using ab26803? Please let us know so that we can cite the reference in this datasheet.

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2 reviews or Q&A

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Votes: Highest First

Immunocytochemistry/ Immunofluorescence abreview for Anti-SAM68 antibody

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat2 cells)
Specification
Rat2 cells
Fixative
Formaldehyde
Permeabilization
Yes - 0.4% triton x-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Oct 22 2012

Western blot abreview for Anti-SAM68 antibody

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Human fibroblast)
Loading amount
20 µg
Specification
Human fibroblast
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris nupage)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
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Verified customer

Submitted Jul 21 2010

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

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