Revision 3

#7558Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, IF-IC, FC-FP

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

160

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9UBN7

Entrez-Gene Id:

10013

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:200 - 1:800
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

HDAC6 (D2E5) Rabbit mAb recognizes endogenous levels of total HDAC6 protein.

Species Reactivity:

Human, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a recombinant protein specific to the carboxy terminus of human HDAC6 protein.

Background

HDAC6 is a class II histone deacetylase enzyme localized to the cytoplasm and associated with the microtubule network (1). It is involved in the regulation of many cellular processes, including cell migration, immune synapse formation, viral infection, and degradation of misfolded proteins (1). HDAC6 contains two tandem catalytic domains that facilitate the deacetylation of multiple protein substrates, including histones and non-histone proteins such as tubulin, cortactin, and HSP90. Despite the ability to deacetylate histone proteins in vitro, there is no evidence for HDAC6-mediated deacetylation of histones in vivo (2,3). The acetylation/deacetylation of tubulin on Lys40 regulates binding and motility of the kinesin-1 motor protein and subsequent transport of cargo proteins such as JNK-interacting protein 1 (JIP1) (4). The acetylation/deacetylation of cortactin regulates cell motility by modulating the binding of cortactin to F-actin (5). Acetylation/deacetylation of HSP90 modulates chaperone complex activity by regulating the binding of an essential cochaperone protein, p23 (6,7). In addition to its role as a protein deacetylase, HDAC6 functions as a component of the aggresome, a proteinaceous inclusion body that forms in response to an accumulation of misfolded or partially denatured proteins (8). Formation of the aggresome is a protective response that sequesters cytotoxic protein aggregates for eventual autophagic clearance from the cell. HDAC6 contains a zinc finger ubiquitin-binding domain that binds both mono- and poly-ubiquitinated proteins (8). HDAC6 binds to both poly-ubiquitinated misfolded proteins and dynein motors, facilitating the transport of misfolded proteins to the aggresome (9,10). HDAC6 is also required for subsequent recruitment of the autophagic machinery and clearance of aggresomes from the cell (11). Thus, HDAC6 plays a key role in the protection against the deleterious effects of pathological protein aggregation that occurs in various diseases, such as neurodegenerative Huntington’s disease (11).

  1. Boyault, C. et al. (2007) Oncogene 26, 5468-76.
  2. Haggarty, S.J. et al. (2003) Proc Natl Acad Sci U S A 100, 4389-94.
  3. Zhang, Y. et al. (2003) EMBO J 22, 1168-79.
  4. Reed, N.A. et al. (2006) Curr Biol 16, 2166-72.
  5. Zhang, X. et al. (2007) Mol Cell 27, 197-213.
  6. Kovacs, J.J. et al. (2005) Mol Cell 18, 601-7.
  7. Murphy, P.J. et al. (2005) J Biol Chem 280, 33792-9.
  8. Seigneurin-Berny, D. et al. (2001) Mol Cell Biol 21, 8035-44.
  9. Kawaguchi, Y. et al. (2003) Cell 115, 727-38.
  10. Boyault, C. et al. (2006) EMBO J 25, 3357-66.
  11. Iwata, A. et al. (2005) J Biol Chem 280, 40282-92.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 3
#7558

HDAC6 (D2E5) Rabbit mAb

Western Blotting Image 1: HDAC6 (D2E5) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using HDAC6 (D2E5) Rabbit mAb.
No image available
Immunohistochemistry Image 1: HDAC6 (D2E5) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using HDAC6 (D2E5) Rabbit mAb.
Immunohistochemistry Image 2: HDAC6 (D2E5) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using HDAC6 (D2E5) Rabbit mAb.
Immunofluorescence Image 1: HDAC6 (D2E5) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of A549 cells, untreated (left) or treated with MG132 (5 μM, 24 hr; right), using HDAC6 (D2E5) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5®#4084 (fluorescent DNA dye).
Flow Cytometry Image 1: HDAC6 (D2E5) Rabbit mAb Expand Image
Flow cytometric analysis of K562 cells using HDAC6 (D2E5) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.