Exalpha Biologicals, Inc.

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FIX&PERM Cell Fixation and Permeabilization Kit

Flow cytometric analyses with monoclonal antibodies were so far mainly restricted to cell surface molecules. Intracellular structures such as cytoplasmic or nuclear enzymes, oncoproteins, cytokines, immunoglobulins etc. were largely excluded from such studies. Also excluded from flow cytometric studies were cytoplasmic localizations of well-established membrane molecules like CD3 and CD22, which, in their cytoplasmic form, are the most reliable lineage markers in undifferentiated leukemia. With the FIX&PERM® Kit flow cytometric analysis of intracellular antigens has become as easy as surface antigen studies. The only prerequisite is the availability of suitable antibody conjugates. Most of the available monoclonal antibody conjugates can be used with the FIX&PERM® Kit, some determinants are sensitive, however, to the fixation step involved. This and the optimal fixation time have to be tested for each reagent.

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Exalpha Biologicals, Inc.

Akt (pan)

  • Product Code: X1890P
  • Size: 200 µl
  • Availability: In Stock In Stock
  • Price (USD): $396

Cat #

X1890P		 Quantity:      

Data Sheet

Product Name

Akt (pan)

Synonyms

Protein Kinase B, RAC-alpha

Host/Source

Rabbit

Isotype

IgG

Product Type

Antigen Immunoaffinity Purified Polyclonal

Reactivity

Human, Mouse .

Applications

Western Blot

Purification

Purified from rabbit serum by sequential epitope-specific chromatography.

Size

200 µl

Price (USD)

$396

Background

Akt, also known as Protein Kinase B (PKB) or RAC- , is a 60 kDa serine/threonine kinase that plays an important role in multiple biological responses including metabolism, cell survival and growth by phosphorylating many proteins including GSK-3 , caspase 9, BAD, and the Forkhead Transcription Factor. Akt is activated by PI3K, which in turn can interact with proteins like FAK thereby linking activation to the cytoskeleton. Akt/PKB is constitutively active in many types of human cancer, and contributes to tumor progression by promoting angiogenesis and cell invasiveness. Akt is fully activated by phosphorylation of threonine 308 by PDK1 and serine 473 by PDK2.

Immunogen

A synthetic peptide derived from the caraboxy-terminus region of human Akt1

Positive Control

Postive Control: 3T3 L1 cell extracts

Formulation

Provided as solution in phosphate buffered saline pH 7.3 with 0.1% BSA, 0.05% sodium azide, and 50% glycerol

Customer Storage

Product should be stored at -20°C. Aliquot to avoid freeze/thaw cycles

Target Molecular Weight

60 kDa

Product Image

Image Legend

Extracts of 3T3L1 cells were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature, then incubated with a 1:1000 (1) or 1:500 (2) dilution of the Akt/PKB pan antibody for two hours at room temperature in a 3% BSA-TBST buffer. After washing, the membrane was incubated with goat F(ab?)2 anti-rabbit IgG HRP-conjugate and signals were detected using the Pierce SuperSignal™ method.

Database Links:

SwissProtP31749Human

References

1. Beffert, U., et al. (2002) Reelin-mediated signaling locally regulates protein kinase B/Akt and glycogen synthase kinase 3beta. J. Biol. Chem. 277(51):49958-49964.
2. Hsu, J., et al. (2001) The AKT kinase is activated in multiple myeloma tumor cells. Blood 98(9):2853-2855.
3. Persad, S., et al. (2001) Regulation of protein kinase B/Akt-threonine-308 phosphorylation by integrin linked kinase (ILK): Critical roles for kinase activity and amino acids arginine-211 and threonine-343. J. Biol. Chem. 276(29):27462-27469.
4. Nelson, J.M. and D.W. Fry. (2001) Akt, MAPK (Erk1/2), and p38 act in concert to promote apoptosis in response to ErbB receptor family inhibition. J. Biol. Chem. 276(18):14842-14847.
5. Genot, E.M., et al. (2000) The T-Cell receptor regulates Akt (Protein Kinase B) via a pathway involving Rac1 and phosphotidylinositide 3-Kinase. Mol. Cell. Biol. 20(15):5469-5478.
6. Datta, S.R., et al. (1997) Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell 91(2):231-241.
7. del Peso, L., et al. (1997) Interleukin-3-induced phosphorylation of BAD through the protein kinase Akt. Science 278(5338):687-689.
8. Dudek, H., et al. (1997) Regulation of neuronal survival by the serine-threonine protein kinase Akt. Science 275(5300):661-665.
9. Franke, T.F., et al. (1997) PI3K: downstream AKTion blocks apoptosis. Cell 88(4):435-437.
10. An, W., et al. (2005) Mechanism of zinc-induced phosphorylation of p70 S6 kinase and glycogen synthase kinase 3 in SH-SY5Y neuroblastoma cells. J. Neurochem. 92:1104-1115 (cites the use of cat. # 44-604G and 44-622G (discontinued).
11. Kumar, P., et al. (2004) p38 MAPK mediates -irradiation-induced endothelial cell apoptosis, and vascular endothelial growth factor protects endothelial cells through the phosphoinositide 3-kinase-Akt-Bcl-2 pathway. J. Biol. Chem. 279(41):43352-43360 (cites the use of cat. # 44-622G (discontinued), 44-680G and 44-684G).