Phospho-PAK (T402) Antibody

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PPS058
Detection of Phospho-PAK (T402) by Western Blot.
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Phospho-PAK (T402) Antibody Summary

Species Reactivity
Human, Mouse, Rat, Primate
Specificity
Human, mouse, non-human primates, and rat ~68-70 kDa PAK-1, 2, 3 phosphorylated at T402. T402 in PAK-2 corresponds to T423 in human PAK-1 in Western blots.
Source
Polyclonal Rabbit IgG
Purification
Antigen Affinity-purified
Immunogen
Phosphopeptide corresponding to amino acid residues surrounding the phospho-T402 of PAK
Formulation
100 μL in 10 mM HEPES (pH 7.5), 150 mM NaCl, 100 μg/mL BSA and 50% glycerol.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1:1000 dilution
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Phospho-PAK (T402) antibody by Western Blot. View Larger

Detection of Phospho-PAK (T402) by Western Blot. Western blot of rat hippocampal lysate showing specific immunolabeling of the approximately 68-70 kDa PAK protein (Control). The phosphospecificity of this labeling is shown in the second lane ( lambda-phosphatase: lambda PPase). The blot is identical to the control except that it was incubated in lambda PPase (1200 units for 30 minutes) before being exposed to the anti-Phospho-PAK-1, 2, 3 (T402). The immunolabeling of PAK is completely eliminated by treatment with lambda PPase.

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
For long-term storage, ≤ -20° C is recommended. Product is stable at ≤ -20° C for at least 1 year.

Background: PAK

PAK (p21-activated kinase) is a general term for a Serine/Threonine kinase that functions downstream of Rho-family GTPases to regulate cytoskeletal (actin) structure and gene transcription. There are two mammalian PAK groups, each with three members. Group I contains PAK-1/ alpha -PAK, PAK-2/ beta -PAK and PAK-3/ gamma -PAK; group II contains PAKs 4-6. All PAKs contain p21-binding domains and a kinase domain. Only group I PAKs are activated by Cdc42 (cell division cycle 24) and exist as dimers. The dimeric form of group I PAKs keep them in an inactive state. This is due to cross-inhibition from one monomer to another. Following p21 binding, these PAKs dissociate and undergo either auto-, or exogenous (PKD1) phosphorylation on a kinase domain Threonine that is embedded in an 18 aa sequence that is absolutely conserved in PAKs 1-3 in both rat and human. In rat, this Threonine occurs at 402 in PAK-2, T422 in PAK-1, and T421 in PAK-3. In human, each position is increased by one (i.e. T403 vs. T402 in rat PAK-2, etc.). Phosphorylation at this homologous site activates the PAKs, which, following additional regulatory phosphorylations, can then interact with desmin, myosin light chain kinase, LIM1, filamin A and/or Merlin. Preparation Prepared from rabbit serum by affinity purification via sequential chromatography

References
  1. Bokoch, G.M. (2003) Annu. Rev. Biochem. 72:743.
  2. Zkumar, R. et al. (2006) Nat. Rev. Cancer 6:459.
  3. Parrini, M.C. et al. (2002) Mol. Cell 9:73.
  4. King, C.C. et al. (2000) J. Biol. Chem. 275:41201.
  5. Chong, C. et al. (2001) J. Biol. Chem. 276:17347.
Long Name
p21/Cdc42/Rac1-activated Kinase 1, 2, 3
Alternate Names
alpha-PAK;IDDMSSD;Non-specific serine/threonine protein kinase;p65-PAK;PAK1;PAKalpha;Serine/threonine-protein kinase PAK 1; PAK

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Phospho-PAK (T402) Antibody
By Anonymous on 11/17/2021
Application: WB Sample Tested: SK-Mel-28 human malignant melanoma cell line Species: Human