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

  • Product Code: X1889M
  • Size: 100 µg
  • Availability: In Stock In Stock
  • Price (USD): $397

Cat #

X1889M		 Quantity:      

Data Sheet

Product Name

Akt

Synonyms

Protein Kinase B, RAC-alpha, EC 2.7.11.1, RAC-PK-alpha, PKB, C-AKT

Host/Source

Mouse

Clone

9Q7

Isotype

IgG3

Product Type

Monoclonal Antibody

Reactivity

Human, Mouse, Rat others not tested

Applications

Western Blot

Purification

Purified from ascites by affinity chromatography.

Size

100 µg

Price (USD)

$397

Background

AKT is a 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 FAK and other cytoskeletal-associated proteins, 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. The reactivity of this antibody with AKT2 and AKT3 has not been tested.

Immunogen

Recombinant human AKT1 protein.

Positive Control

Positive Control: Human Jurkat cells, mouse 3T3-L1 cells, and rat PC-12 cells. Found in all human cell types so far analyzed. Found in the cytoplasm (and in the nucleus after activation by integrin-linked protein kinase 1)

Formulation

Provided as solution in phosphate buffered saline pH 7.2 with 0.1% BSA and 0.01% sodium azide

Customer Storage

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

Target Molecular Weight

60 kDa

Product Image

Image Legend

Proteins from cell extracts of human Jurkat cells (lane 1), mouse 3T3-L1 cells (lane 2), and rat PC-12 cells (lane 3) were resolved by SDS-PAGE and transferred to PVDF. The membranes were incubated with this AKT monoclonal antibody (clone 9Q7) at a concentration of 1 µg/mL for two hours at room temperature. After washing, the membranes were incubated with a goat F(ab?)2 anti-mouse IgG alkaline phosphatase conjugated antibody (Cat. # AMI4405) at a 1:2000 dilution. Bands were detected with CDP-substrate using the WesternStarTM method (Tropix) and Kodak BioMax film.

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 phosphatidylinositide 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 AKT phosphorylation blocks apoptosis. Cell 88(4):435-437.