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O této položce
UNSPSC Code:
12352203
NACRES:
NA.41
eCl@ss:
32160702
Clone:
6-11B-1, monoclonal
Species reactivity:
sea urchin, bovine, monkey, Chlamydomonas, Drosophila, mouse, rat-kangaroo, human
Application:
DB, EM, ICC, WB
Citations:
40
Technický servis
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mouse
Quality Segment
antibody form
purified immunoglobulin
antibody product type
primary antibodies
clone
6-11B-1, monoclonal
species reactivity
sea urchin, bovine, monkey, Chlamydomonas, Drosophila, mouse, rat-kangaroo, human
technique(s)
dot blot: suitable, electron microscopy: suitable, immunocytochemistry: suitable, western blot: suitable
isotype
IgG2bκ
GenBank accession no.
UniProt accession no.
shipped in
wet ice
target post-translational modification
acetylation (Lys40)
Gene Information
human ... TUBA1A(7846)
General description
Microtubules are cytoskeletal filaments composed of of alpha-/beta-tubulin heterodimers that self-assemble head-to-tail to form protofilaments and laterally to form a hollow tube. Various post-translational modifications (PTMs), including acetylation, polyglutamylation, polyglycylation, detyrosination, phosphorylation, and palmitoylation, can regulate the polymerization properties of tubulins and/or their interactions with microtubule associated proteins (MAPs) and motor proteins. A subset of cytoplasmic microtubules and microtubules in the spindle, axon, and cilia are known to contain alpha-tubulin acetylated via the epsilon-amino group of Lysine-40 (K40). The GNAT lysine acetyltransferase family member MEC-17 (or alpha-tubulin acetyltransferase/alphaTAT) catalyzes K40 acetylation, while HDAC6 and sirtuin2 (SIRT2) are known to catalyze tubulin deacetylation. Experimental evidences indicate that K40 acetylation primes alpha-tubulin for further PTM(s), which cannot be reversed by later deactylation of K40ac. Subsequently, the acetylated and deacetylated alpha-tubulin in native microtubules are structurally distinct from that of unacetylated alpha-tubulin (never before has its K40 acetylated).
~50 kDa observed
Immunogen
15 S dynein fraction from sea urchin sperm axoneme.
Application
Research Category
Cell Structure
Cell Structure
Research Sub Category
Adhesion (CAMs)
Adhesion (CAMs)
This Anti-acetyl-alpha tubulin Antibody, clone 6-11B-1 is validated for use in Western Blotting, Immunocytochemistry, Dot Blot and Electron Microscopy for the detection of acetyl-alpha tubulin.
Western Blotting Analysis: 4.0 µg/mL from a representative lot detected acetyl-alpha tubulin in 10 µg of mouse brain tissue lysate.
Immunocytochemistry Analysis: A representative lot detected acetylated alpha-tubulin in cultured normal human kidney epithelial cells (Courtesy of Dr Christopher Ward, University of Kansas Medical Center).
Western Blotting Analysis: A representative lot detected alpha-tubulin in sea urchin sperm axoneme lysates, but not in lysates from taxol-stabilized microtubules from sea urchin eggs (Piperno, G., and Fuller, M. (1985). J Cell Biol.101(6):2085-2094).
Dot Blot Analysis: A representative lot detected alpha-tubulin in both the ionic detergent Sarkosyl-soluble and insoluble fractions from sea urchin spern axoneme extracts (Piperno, G., and Fuller, M. (1985). J Cell Biol.101(6):2085-2094).
Immunocytochemistry Analysis: A representative lot detected acetylated and deacetylated, but not unacetylated alpha-tubulin in COS-7 green monkey kidney fibroblasts and Ptk2 rat kangaroo kidney epithelial cells by fluorescent immunocytochemistry (Soppina V., et al. (2012). PLoS One. 7(10):e48204).
Electron Microscopy Analysis: The Fab fragment of clone 6-11B-1 has been shown to stain Taxol-stabilized microtubules polymerized in vitro from purified bovine brain tubulin with or without SIRT2-catalyzed deacetylation (Soppina V., et al. (2012). PLoS One. 7(10):e48204).
Immunocytochemistry Analysis: A representative lot detected acetylated alpha-tubulin in cultured normal human kidney epithelial cells (Courtesy of Dr Christopher Ward, University of Kansas Medical Center).
Western Blotting Analysis: A representative lot detected alpha-tubulin in sea urchin sperm axoneme lysates, but not in lysates from taxol-stabilized microtubules from sea urchin eggs (Piperno, G., and Fuller, M. (1985). J Cell Biol.101(6):2085-2094).
Dot Blot Analysis: A representative lot detected alpha-tubulin in both the ionic detergent Sarkosyl-soluble and insoluble fractions from sea urchin spern axoneme extracts (Piperno, G., and Fuller, M. (1985). J Cell Biol.101(6):2085-2094).
Immunocytochemistry Analysis: A representative lot detected acetylated and deacetylated, but not unacetylated alpha-tubulin in COS-7 green monkey kidney fibroblasts and Ptk2 rat kangaroo kidney epithelial cells by fluorescent immunocytochemistry (Soppina V., et al. (2012). PLoS One. 7(10):e48204).
Electron Microscopy Analysis: The Fab fragment of clone 6-11B-1 has been shown to stain Taxol-stabilized microtubules polymerized in vitro from purified bovine brain tubulin with or without SIRT2-catalyzed deacetylation (Soppina V., et al. (2012). PLoS One. 7(10):e48204).
Biochem/physiol Actions
Experimental evidences indicate that the acetylated and deacetylated α-tubulin in native microtubules are structurally distinct from that of unacetylated α-tubulin, and that clone 6-11B-1 recognizes acetylated and deacetylated α-tubulin, but not unacetylated (never acetylated) α-tubulin. While 6-11B-1 immunostaining pattern resembles that of acetyl-Lys40 antibodies on normal cylcling cells, caution must be taken when interpreting immunostaining results using this mAb on cells subjected to tubulin deacetylation manipulations by exogenous deacetylase expression or via chemical means.
Physical form
Format: Purified
Protein G Purified
Purified mouse monoclonal IgG2bκ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Preparation Note
Stable for 1 year at 2-8°C from date of receipt.
Analysis Note
Evaluated by Western Blotting in HeLa cell lysate.
Western Blotting Analysis: 4.0 µg/mL of this antibody detected acetyl-alpha tubulin in 10 µg of HeLa cell lysate.
Western Blotting Analysis: 4.0 µg/mL of this antibody detected acetyl-alpha tubulin in 10 µg of HeLa cell lysate.
Other Notes
Concentration: Please refer to lot specific datasheet.
Legal Information
GenBank is a registered trademark of United States Department of Health and Human Services
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.
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Skladovací třída
12 - Non Combustible Liquids
wgk
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
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