Cell Signaling Technology

Product Pathways - Apoptosis

Mcl-1 (D2W9E) Rabbit mAb #94296

sc-819  

No. Size Price
94296S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
94296 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat, Endogenous 40 (human), 35 (mouse) Rabbit IgG
IP 1:100
F 1:50
IF-IC 1:800

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),

Specificity / Sensitivity

Mcl-1 (D2W9E) Rabbit mAb recognizes endogenous levels of total Mcl-1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro60 of mouse Mcl-1 protein.

IF-IC

IF-IC

Confocal immunofluorescent analysis of MCF7 (left), L-929 (center), and SK-OV-3 (right) cells using Mcl-1 (D2W9E) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Mcl-1 (D2W9E) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

IP

IP

Immunoprecipitation of Mcl-1 from MCF7 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Mcl-1 (D2W9E) Rabbit mAb. Western blot analysis was performed using Mcl-1 (D2W9E) Rabbit mAb. A conformation-specific secondary antibody was used to avoid cross-reactivity with IgG.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected or transfected with a construct expressing full-length human Mcl-1 (hMcl-1; +) or mouse Mcl-1 (mMcl-1; +), using Mcl-1 (D2W9E) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of SK-OV-3 cells (blue) and MCF7 cells (green) using Mcl-1 (D2W9E) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of L-929 cells using Mcl-1 (D2W9E) Rabbit mAb (blue) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was use as a secondary antibody.

Background

Mcl-1 is an anti-apoptotic member of the Bcl-2 family originally isolated from the ML-1 human myeloid leukemia cell line during phorbol ester-induced differentiation along the monocyte/macrophage pathway (1). Similar to other Bcl-2 family members, Mcl-1 localizes to the mitochondria (2), interacts with and antagonizes pro-apoptotic Bcl-2 family members (3), and inhibits apoptosis induced by a number of cytotoxic stimuli (4). Mcl-1 differs from its other family members in its regulation at both the transcriptional and post-translational level. First, Mcl-1 has an extended amino-terminal PEST region, which is responsible for its relatively short half-life (1,2). Second, unlike other family members, Mcl-1 is rapidly transcribed via a PI3K/Akt dependent pathway, resulting in its increased expression during myeloid differentiation and cytokine stimulation (1,5-7). Mcl-1 is phosphorylated in response to treatment with phorbol ester, microtubule-damaging agents, oxidative stress, and cytokine withdrawal (8-11). Phosphorylation at Thr163, the conserved MAP kinase/ERK site located within the PEST region, slows Mcl-1 protein turnover (10) but may prime the GSK-3 mediated phosphorylation at Ser159 that leads to Mcl-1 destabilization (11). Mcl-1 deficiency in mice results in peri-implantation lethality (12). In addition, conditional disruption of the corresponding mcl-1 gene shows that Mcl-1 plays an important role in early lymphoid development and in the maintenance of mature lymphocytes (13).

  1. Kozopas, K.M. et al. (1993) Proc Natl Acad Sci USA 90, 3516-20.
  2. Yang, T. et al. (1995) J Cell Biol 128, 1173-84.
  3. Sato, T. et al. (1994) Proc Natl Acad Sci USA 91, 9238-42.
  4. Zhou, P. et al. (1997) Blood 89, 630-43.
  5. Wang, J.M. et al. (1999) Mol Cell Biol 19, 6195-206.
  6. Jourdan, M. et al. (2003) Oncogene 22, 2950-9.
  7. Chao, J.R. et al. (1998) Mol Cell Biol 18, 4883-98.
  8. Domina, A.M. et al. (2000) J Biol Chem 275, 21688-94.
  9. Inoshita, S. et al. (2002) J Biol Chem 277, 43730-4.
  10. Domina, A.M. et al. (2004) Oncogene 23, 5301-15.
  11. Maurer, U. et al. (2006) Mol Cell 21, 749-60.
  12. Rinkenberger, J.L. et al. (2000) Genes Dev 14, 23-7.
  13. Opferman, J.T. et al. (2003) Nature 426, 671-6.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

Companion Products


For Research Use Only. Not For Use In Diagnostic Procedures.

The Alexa Fluor dye antibody conjugates in this product are sold under license from Life Technologies Corporation for research use only, except for use in combination with DNA microarrays. The Alexa Fluor® dyes (except for Alexa Fluor® 430 dye) are covered by pending and issued patents. Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

DRAQ5 is a registered trademark of Biostatus Limited.

DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

Tween is a registered trademark of ICI Americas, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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