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Anti-KCC2 (SLC12A5) Antibody

K+-Cl- cotransporter 2, Electroneutral potassium-chloride cotransporter 2, Neuronal K-Cl cotransporter, Solute carrier family 12 member 5

Cat #: AKT-005
Alternative Name K+-Cl- cotransporter 2, Electroneutral potassium-chloride cotransporter 2, Neuronal K-Cl cotransporter, Solute carrier family 12 member 5
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)KNPANTRLRLNVPE, corresponding to amino acid residues 967-980 of rat KCC2 (Accession Q63633). Intracellular, C-terminus.
Accession (Uniprot) Number Q63633
Gene ID 171373
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Human – identical; mouse – 13/14 amino acid residues identical.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
Standard quality control of each lot Western blot analysis.
Applications: if, ih, wb
May also work in: ifc*, ip*
Western blot
  • Western blot analysis of rat brain membranes:
    Western blot analysis of rat brain membranes:
    1. Anti-KCC2 (SLC12A5) Antibody (#AKT-005), (1:500).
    2. Anti-KCC2 (SLC12A5) Antibody, preincubated with KCC2/SLC12A5 Blocking Peptide (#BLP-KT005).
  • Western blot analysis of mouse brain lysates:
    Western blot analysis of mouse brain lysates:
    1. Anti-KCC2 (SLC12A5) Antibody (#AKT-005), (1:200).
    2. Anti-KCC2 (SLC12A5) Antibody, preincubated with KCC2/SLC12A5 Blocking Peptide (#BLP-KT005).
Immunohistochemistry
  • Expression of KCC2 in rat cerebellum and hippocampus.
    Expression of KCC2 in rat cerebellum and hippocampus.
    Immunohistochemical staining of perfusion-fixed frozen rat brain sections with Anti-KCC2 (SLC12A5) Antibody (#AKT-005), (1:1000), followed by goat anti-rabbit-AlexaFluor-488. A. KCC2 immunoreactivity in the cerebellum (green) is detected in the granule layer (GL) and molecular layer (MOL). B. KCC2 staining in the hippocampal dentate gyrus region (green) is observed in the outer molecular layer (OML) and the hilus (H). Cell nuclei are stained with DAPI (blue).
References
  1. Côme, E. et al. (2019) Front. Cell. Neurosci. 13, 48.
  2. Fukuda, A. and Watanabe, M. et al. (2019) Brain Res. 1710, 1.
Scientific background

KCC2 (K+-Cl-cotransporter 2) is a glycoprotein neuron-specific cotransporter and a member of the cation-chloride co-transporter (CCC) family encoded by the Slc12a5 gene. KCC2 is responsible for maintaining low Cl levels in neurons using the potassium gradient. In addition, KCC2 regulates the formation, functional maintenance and plasticity of glutamatergic synapses. The proper inhibitory functions of GABA are maintained by KCC2 since GABA is dependent on the Cl driving force promoted by the intracellular concentration of Cl 1,2.

KCC2 has a predicted structure of 12 transmembrane segments, six extracellular loops flanked by a short intracellular amino terminal and a long intracellular carboxy-terminal1,2.

KCC2 protein expression is detected in the central nervous system including the spinal cord, thalamus, cerebellum, hippocampus, cortical structures and the auditory brainstem1.

Human KCC2 mutations are associated with severe early-onset epileptic encephalopathy and febrile seizures1,2.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat
Image & Title:


Multiplex staining of KCC2 and GABA(A) α1 Receptor in rat cerebellum.Immunohistochemical staining of perfusion-fixed frozen rat brain sections using rabbit Anti-KCC2 (SLC12A5) Antibody (#AKT-005), (1:400) and Guinea pig Anti-GABA(A) α1 Receptor (extracellular) Antibody (#AGP-083), (1:400). A. KCC2 immunoreactivity (green) is detected in both the granule layer (GL) and the molecular layer (MOL). B. GABA(A) α1 receptor staining (red) is observed mainly in the granule layer (GL). C. Merge of the two images demonstrates colocalization in the granule layer (arrows). Cell nuclei are stained with DAPI (blue).

Last update: 12/08/2021

Anti-KCC2 (SLC12A5) Antibody (#AKT-005) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize KCC2 from human, rat, and mouse samples.

For research purposes only, not for human use
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