07-415-I

Anti-Sam 68 Antibody

serum, from rabbit

• Synonym(s): KH domain-containing, RNA-binding, signal transduction-associated protein 1, GAP-associated tyrosine phosphoprotein p62, Src-associated in mitosis 68 kDa protein, Sam68, p21 Ras GTPase-activating protein-associated p62, p68
• UNSPSC Code: 12352203
• eCl@ss: 32160702
• NACRES: NA.41

Properties

• biological source: rabbit
• Quality Level: 100
• antibody form: serum
• antibody product type: primary antibodies
• clone: polyclonal
• species reactivity: human, mouse
• species reactivity (predicted by homology): rat (based on 100% sequence homology)
• technique(s): immunocytochemistry: suitable; immunohistochemistry: suitable; immunoprecipitation (IP): suitable; western blot: suitable
• NCBI accession no.: NP_035447
• UniProt accession no.: Q60749
• shipped in: wet ice
• target post-translational modification: unmodified
• Gene Information: human ... KHDRBS1(10657) ; mouse ... Khdrbs1(20218) ; rat ... Khdrbs1(117268)

Description

General description

KH domain-containing, RNA-binding, signal transduction-associated protein 1 (UniProt Q60749; also known as GAP-associated tyrosine phosphoprotein p62, Khdrbs1,p68, p21 Ras GTPase-activating protein-associated p62, Sam68, Src-associated in mitosis 68 kDa protein) is encoded by the Khdrbs1 gene (Gene ID 20218) in murine species. Sam68 is a member of the “signal transduction and activation of RNA” (STAR) family of KH domain-containing RNA-binding proteins that mediates pre-mRNA splicing, mRNA export, and protein translation. It binds to UAAA, UUUA, and UUUUUU motifs with high affinity. RNA immunoprecipitation (RIP) techniques using lysates from UV cross-linked NIH 3T3 cells have identified 23 Sam68 complex-associated mRNA species, including Thrap1, bri3bp, Myl6, Copeb/KLF6/Zf9, Hmgn1, Smc6, and biglycan. In addition, Sam68 is found to be a key regulator of Nrxn1 alternative splicing in the central nervous system in a neuronal activity-dependent manner. Sam68 is phosphorylated on three tyrosine residues (pTyr435, pTyr440, and pTyr443) within the C-terminal nuclear localization signal (NLS) region by the breast tumor kinase (BRK) following EGF stimulation of breast tumor cells with pTyr440 being a principal modulator of Sam68 nuclear localization. Studies comparing wild-type and Sam68-/- knockout mice revealed a positive regulatory role of Sam68 toward adipocyte differentiation, as well as a negative impact of Sam68 against osteoblast differentiation among aged mice.

Specificity

This antiserum (also known as AD-1 or AD1) recognizes Sam68, but not two other GSG (GRP33, Sam68, GLD-1) proteins, the Sam68-like mammalian proteins SLM-1 and SLM-2.

Immunogen

Epitope: aa 331-348

KLH-conjugated peptide corresponding to amino acid residues 331-348 of human Sam 68

Application

Research Category
Signaling

Research Sub Category
Signaling Neuroscience

This Anti-Sam 68 Antibody is validated for use in Western Blotting, Immunohistochemistry, Immunocytochemistry and Immunoprecipitation for the detection of Sam 68 .

Western Blotting Analysis: 1:500-1:10,000 of a representative lot detected Sam 68 in RIPA lysates from HeLa cells.
Immunohistochemistry Analysis: A 1:600 dilution from a representative lot detected Sam 68 in mouse colon tissue (Data courtesy of Stephane Richard, McGill University).
Western Blotting Analysis: A representative lot detected Sam 68 in endogenous Sam 68 and exogenously expressed GFP-Sam 68 fusion in HeLa cells (Chen, T., et al. (1999). Mol Biol Cell. 10(9):3015-3033).
Western Blotting Analysis: A representative lot detected Sam 68 in HEK293 transfected with GFP-Sam68, mouse cortex, hippocampus, cerebellum, and mouse brain (Iijima, T., et al. (2011). Cell. 147(7):1601-1614).
Western Blotting Analysis: A representative lot detected Sam 68 in HeLa cells transfected with GFP-Sam68, MDA-MB-231, MDA-MB-468, and BT20 (Lukong, K.E., et al. (2005). J Biol Chem. 280(46):38639-38647).
Western Blotting Analysis: A representative lot detected Sam 68 in lysates from isolated mouse cells and mouse embryonic cell (Richard, S., et al. (2005). PLoS Genet. 1(6):e47).
Immunohistochemistry Analysis: A representative lot detected Sam 68 in mouse cerebellum, parasagittal section of brain stem and cerebellum (Iijima, T., et al. (2011). Cell. 147(7):1601-1614).
Immunohistochemistry Analysis: A representative lot detected Sam 68 in embryonic mice (Richard, S., et al. (2005). PLoS Genet. 1(6):e47).
Immunocytochemistry Analysis: A representative lot detected Sam 68 in HeLa cells (Chen, T., et al. (1999). Mol Biol Cell. 10(9):3015-3033).
Immunoprecipitation Analysis: A representative lot immunoprecipitated Sam 68 in HeLa, HeLa cells transfected with GFP-Sam68, and MDA-MB-468 (Lukong, K.E., et al. (2005). J Biol Chem. 280(46):38639-38647).

Quality

Evaluated by Western Blotting in MDA-MB-468 cell lysate.

Western Blotting Analysis: A 1:500 dilution of this antibody detected Sam68 in 10 µg of MDA-MB-468 cell lysate.

Target description

~68 kDa observed

Linkage

Replaces: 07-415

Physical form

Rabbit polyclonal serum with 0.05% sodium azide.

Unpurified

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 1