- Datasheet
- Specific References (1)
- Protocols
Overview
- Product nameAnti-SAM68 antibody
See all SAM68 primary antibodies - DescriptionRabbit polyclonal to SAM68
- Tested applicationsSuitable for: ICC/IF, WB, IHC-Pmore details
- Species reactivityReacts 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.
(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
- FormLiquid
- Storage instructionsShipped 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 bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- ClonalityPolyclonal
- IsotypeIgG
- Research areas
Associated products
- Compatible Secondaries
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
- FunctionRecruited 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 specificityUbiquitously 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 similaritiesBelongs to the KHDRBS family.
Contains 1 KH domain. - Developmental stageIsoform 3 is only expressed in growth-arrested cells.
- DomainThe 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
modificationsTyrosine 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 localizationNucleus. Membrane.
- Information by UniProt
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Database links
- Entrez Gene: 374071 Chicken
- Entrez Gene: 10657 Human
- Entrez Gene: 20218 Mouse
- Entrez Gene: 117268 Rat
- Omim: 602489 Human
- SwissProt: Q8UUW7 Chicken
- SwissProt: Q07666 Human
- SwissProt: Q3U8T3 Mouse
see all -
Alternative names
- FLJ34027 antibody
- GAP associated tyrosine phosphoprotein p62 antibody
- GAP-associated tyrosine phosphoprotein p62 antibody
see all
Anti-SAM68 antibody images
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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) 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.
Lysates/proteins at 20 µg per lane.
Secondary
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?) -
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
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.
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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.
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Western blot - Anti-SAM68 antibody (ab26803)This image is courtesy of an anonymous AbreviewAnti-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 minuteThis image is courtesy of an anonymous Abreview
Blocking Step: 5% BSA for 1 hour at 25°C -
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 for Anti-SAM68 antibody (ab26803)
This product has been referenced in:
- 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) »
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"