Rabbit Potassium Chloride Cotransporter (KCC2) Polyclonal Antibody | anti-KCC2 antibody

Anti-Phospho-Thr1007 Potassium Chloride Cotransporter (KCC2) Antibody

Cat. Num. AAA500088

• Gene Names: Slc12a5; Kcc2
• Reactivity: Mouse, Rat; Predicted: Bovine, Human, Non-human primate, Pig, Sheep
• Applications: Western Blot
• Purity: Prepared from pooled rabbit serum by affinity purification via sequential chromatography on phospho and non-phosphopeptide affinity columns.
• Synonyms: Potassium Chloride Cotransporter (KCC2); Polyclonal Antibody; Anti-Phospho-Thr1007 Potassium Chloride Cotransporter (KCC2) Antibody; anti-KCC2 antibody

• Host: Rabbit
• Reactivity: Mouse, Rat; Predicted: Bovine, Human, Non-human primate, Pig, Sheep
• Clonality: Polyclonal
• Specificity: Specific for endogenous levels of the ~135 kDa KCC2 protein phosphorylated at Thr1007.; Immunolabeling is completely eliminated by treatment with Lambda-Ptase.
• Purity/Purification: Prepared from pooled rabbit serum by affinity purification via sequential chromatography on phospho and non-phosphopeptide affinity columns.
• Form/Format: 100 ul in 10 mM HEPES (pH 7.5), 150 mM NaCl, 100 ug per ml BSA and 50% glycerol.
• Sequence Length: 1116

• Applicable Applications for anti-KCC2 antibody: Western Blot (WB)
• Application Notes: WB: 1:1000
• Immunogen Species: Rat
• Immunogen Information: Synthetic phospho-peptide corresponding to amino acid residues surrounding Ser940 conjugated to KLH.
• Antigen: Phosphopeptide corresponding to amino acid residues surrounding the phospho-Thr 1007 of mouse KCC2.
• Quality Control Tests: Western blots performed on each lot.
• Preparation and Storage: Storage at -20 degree C is recommended, as aliquots may be taken without freeze/thawing due to presence of 50% glycerol. Stable for at least 1 year at -20 degree C.

Western Blot (WB)

(Western blot of rat hippocampal lysate showing specific labeling of the ~135 kDa KCC2 protein phosphorylated at Thr1007 in the first lane (-). Phosphospecificity is shown in the second lane (+) where immunolabeling is completely eliminated by blot treatment with lambda phosphatase (lambda-Ptase, 1200 units for 30 min).)

Related Product Information for anti-KCC2 antibody

KCC2 is widely thought to be expressed exclusively in neurons where it is responsible for maintaining low intracellular chloride concentration to drive hyperpolarizing post-synaptic responses to the inhibitory neurotransmitters GABA and glycine (Lee et al., 2007). KCC2 is expressed in most adult neurons, and expression levels correlate well with the maturation state of neurons. N-ethylmaleimide (NEM) has been show to increase phosphorylation of the Ser940 residue, while decreasing phosphorylation of T1007 residue. Dephosphorylation of residues T906 and T1007 correlates with increased KCC2 activity (Moss et al., 2017).

References

1. Conway, L.C., Cardarelli, R.A., Moore, Y.E., Jones, K., McWilliams, L.J., Baker, D.J., Burnham, M.P., Bürli, R.W., Wang, Q., Brandon, N.J. and Moss, S.J., 2017. N-ethylmaleimide increases KCC2 activity by modulating transporter phosphorylation. Journal of Biological Chemistry, pp.jbc-M117. 2. Lee HH, Walker JA, Williams JR, Goodier RJ, Payne JA, Moss SJ (2007) Direct protein kinase C-dependent phosphorylation regulates the cell surface stability and activity of the potassium chloride cotransporter KCC2. J Biol Chem. 2007 Oct 12; 282(41):29777-84.

NCBI and Uniprot Product Information

• NCBI GI #: 19705463
• NCBI GeneID: 171373
• NCBI Accession #: NP_599190.1
• NCBI GenBank Nucleotide #: NM_134363.1
• UniProt Accession #: Q63633
• Molecular Weight: ~135 kDa
• NCBI Official Full Name: solute carrier family 12 member 5
• NCBI Official Synonym Full Names: solute carrier family 12 member 5
• NCBI Official Symbol: Slc12a5
• NCBI Official Synonym Symbols: Kcc2
• NCBI Protein Information: solute carrier family 12 member 5
• UniProt Protein Name: Solute carrier family 12 member 5
• UniProt Gene Name: Slc12a5
• UniProt Synonym Gene Names: Kcc2; rKCC2
• UniProt Entry Name: S12A5_RAT

NCBI Description

neuronal specific potassium-chloride cotransporter isoform; may play a role in response to neuronal injury [RGD, Feb 2006]

Uniprot Description

Function: Mediates electroneutral potassium-chloride cotransport in mature neurons. Transport occurs under isotonic conditions, but is activated 20-fold by cell swelling. Important for Cl- homeostasis in neurons. Necessary for the ontogenic change in response to gamma-aminobutyric acid (GABA) from depolarization to hyperpolarization during neuronal development. Ref.3

Enzyme regulation: Inhibited by WNK3

By similarity.

Subunit structure: Homomultimer and heteromultimer with other K-Cl cotransporters. Interacts with AP2A1

By similarity. Ref.4

Subcellular location: Membrane; Multi-pass membrane protein.

Tissue specificity: Highly expressed in brain. Not detected in other tissues. Highly expressed in pyramidal neurons and in neurons throughout the cortex, hippocampus, the granular layer of the cerebellum and in groups of neurons throughout the brainstem. Barely detectable in dorsal-root ganglions.

Developmental stage: Detected in thalamus, but not in hippocampus and neocortex at E20. At birth barely detectable in hippocampus. Expression increases steeply from day 5 to day 9 and then stabilizes at adult levels.

Sequence similarities: Belongs to the SLC12A transporter family.