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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We are pleased to announce that Peter Rutten has now started in his role as Operations Director here at Exalpha Biologicals Inc. Peter has a Master’s of Science degree in Industrial and Organizational Psychology and this has lead him down a very business orientated career path. Peter is looking forward to working with the team at Exalpha. Peters primary focus is the customer experience and he will be working with the Laboratory team, the Quality Control team and the Order Processing team to ensure this focus is achieved. We all wish Peter well in his new role.

Exalpha Biologicals, Inc.

phospho Akt/PKB [pS473]

  • Product Code: X2010M
  • Size: 10 Miniblots
  • Availability: In Stock In Stock
  • Price (USD): $562

Cat #

X2010M		 Quantity:      

Data Sheet

Product Name

phospho Akt/PKB [pS473]

Synonyms

Protein Kinase B or RAC-alpha

Host/Source

Rabbit

Clone

14-6

Isotype

IgG

Product Type

Phosphorylation Site-Specific Monoclonal Antibody

Reactivity

Human, Mouse, Rat

Applications

Western Blot, Immuncytochemistry,

Purification

Protein A/G Chromatography

Size

10 Miniblots

Price (USD)

$562

Background

Akt, also known as Protein Kinase B (PKB) or RAC- , is a 65 kDa serine/threonine kinase that plays an important role in diverse biological responses such as regulation of 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 such as FAK (when FAK is phosphorylated at tyrosine 397) thereby linking activation to the cytoskeleton. Akt is phosphorylated on threonine 308 by PDK1 and on serine 473 by PDK2. Phosphorylation at serine 473 is required for full activation of Akt.

Immunogen

Chemically synthesized phosphopeptide derived from the region of human Akt that contains serine 473.

Positive Control

Mouse NIH3T3 cells stimulated with PDGF; 3T3-L1 adipocytes stimulated with OSM and IFN- .

Formulation

Provided as solution in phosphate buffered saline, pH 7.3, with 1.0 mg/ml 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

65 kDa

Product Image

Image Legend

Immunofluorescence Staining Serum-starved NIH3T3 cells left untreated (left) or treated with PDGF (right) were fixed prior to immunostaining with the Akt [pS473] rabbit monoclonal antibody. The signal was detected with an anti-rabbit FITC conjugated secondary antibody. The data show that the antibody detected phosphorylated Akt in PDGF-treated NIH3T3 cells. Cells were counterstained with DAPI. Please visit our website (biosource.com) to view the images in full color.

Database Links:

SwissProtP47196Rat
SwissProtP31750Mouse
SwissProtP31749Human

References

1. 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).
2. 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, 44-680G and 44-684G).
3. Kovalenko, D., et al. (2003) Sef inhibits fibroblast growth factor signaling by inhibiting FGFR1 tyrosine phosphorylation and subsequent ERK activation. J. Biol. Chem. 278(16):14087-14091 (cites the use of cat. # 44-622G).
4. Yu, D., et al. (2003) Redundancy of radioresistant signaling pathways originating from insulin-like growth factor I receptor. J. Biol. Chem. 278(9):6702-6709 (cites the use of cat. # 44-622G).
5. 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 (cites the use of cat. # 44-622G).
6. Morel, J.C., et al. (2002) Signal transduction pathways involved in rheumatoid arthritis synovial fibroblast interleukin-18-induced vascular cell adhesion molecule-1 expression. J. Biol. Chem. 277(38):34679-34691 (cites the use of cat. # 44-506G, 44-622G and 44-660G).
7. Welch, S., et al. (2002) Cardiac-specific IGF-1 expression attenuates diluted cardiomyopathy in tropomodulin-overexpressing transgenic mice. Circ. Res. (90):641-648 (cites the use of cat. # 44-622G).