Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Ca / cAMP / Lipid Signaling

Phospho-MARCKS (Ser167/170) (D13E4) XP® Rabbit mAb #8722

Applications Reactivity Sensitivity MW (kDa) Isotype
W IF-IC F H M R (C) (B) Endogenous 75 rodent, 80 human Rabbit IgG

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  C=Chicken  B=Bovine
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-MARCKS (Ser167/170) (D13E4) XP® Rabbit mAb recognizes endogenous levels of MARCKS protein only when phosphorylated at Ser167 and Ser170. This antibody may also detect MARCKS mono-phosphorylated at Ser167.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser167 and Ser170 of human MARCKS protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from serum-starved SH-SY5Y and SK-N-MC cells, untreated (-) or treated with TPA #4174 (200 nM, 15 min; +), using Phospho-MARCKS (Ser167/170) (D13E4) XP® Rabbit mAb (upper) and MARCKS (D88D11) XP® Rabbit mAb #5607 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines, serum-starved overnight and then untreated (-) or treated with TPA #4174 (200 nM, 15 min; +), using Phospho-MARCKS (Ser167/Ser170) (D13E4) XP® Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, serum-starved (left), treated with TPA #4174 (200 nM, 15 min; middle) or treated with TPA #4174 and λ phosphatase (right), using Phospho-MARCKS (Ser167/170) (D13E4) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


Background

Myristoylated Alanine-Rich C-Kinase Substrate (MARCKS) is a major PKC substrate expressed in many cell types. MARCKS has been implicated in cell motility, cell adhesion, phagocytosis, membrane traffic, and mitogenesis (1). PKC phosphorylates Ser159, 163, 167, and 170 of MARCKS in response to growth factors and oxidative stress. Phosphorylation at these sites regulates the calcium/calmodulin binding and filamentous (F)-actin cross-linking activities of MARCKS (2-4). Phosphorylation by PKC also results in translocation of MARCKS from the plasma membrane to the cytoplasm (5).

  1. Ramsden, J.J. (2000) Int J Biochem Cell Biol 32, 475-9.
  2. Heemskerk, F.M. et al. (1993) Biochem Biophys Res Commun 190, 236-41.
  3. Graff, J.M. et al. (1989) J Biol Chem 264, 21818-23.
  4. Hartwig, J.H. et al. (1992) Nature 356, 618-22.
  5. Thelen, M. et al. (1991) Nature 351, 320-2.

Application References

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Companion Products

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

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