EnCor Biotechnology

Mouse Monoclonal Antibody to Phospho-MAP-τ-Tp205 (phospho-MAPT T205), Cat# MCA-1H103

$250.00
Description

      The MCA-1H103 antibody was raised against amino acids 190-215 of low molecular weight four tubulin binding repeat τ (aka MAP-tau). This peptide included to known phosphorylation sites phospho-Ser202 and phospho-Thr205 using the sequence in NP_005901.2. This peptide sequence is expressed in all known human τ isotypes and is totally conserved in all mammals, for exact sequence see τ-Epitopes.pdf. The antibody recognizes S202/T205 phosphorylated tau and also T205 phosphoylated tau, but shows no reactivity with non phosphorylated τ only phosphorylated on S202. We have two other mouse monoclonal antibodies binding to other defined epitopes on human τ, MCA-5B10 and MCA-2E9. Both antibodies recognize the unphosphorylated forms of τ and like MCA-1H103 work well on western blots, IF, ICC and IHC.

 Collections:
Amount: 100µL of 1mg/mL
Amount: 100µL of 1mg/mL
Mixed culture from E18 embryonic rat cortex grown for 7 days in Vitro and stained with 1:1,000 MCA-1H103 in green and 1:10,000 rabbit polyclonal antibody to MAP2AB RPCA-MAP2AB in red. DNA in nuclei is revealed with DAPI stain in blue. MCA-IH103 strongly stains the perikarya, dendrites and axons of developing neurons anastomosing throughout the culture. The MAP2 antibody binds only the high molecular forms of MAP2 which are expressed only in dendrites and also later in neuronal development. As a result this antibody recognizes a few dendritic profiles in certain developing neurons. The inset at top right shows only the MAP2AB signal in the region delineated by the box close to the center of the image.
Western blot analysis of blot using mouse mAb to phospho-tau, MCA-1H103, dilution 1:2,000 in green: [1] protein standard (red), [2] human recombinant protein Map-tau, (1ug), [3] rat brain, and [4] embryonic cortical neuron cell culture lysates. MCA-1H103 antibody raised against a peptide including phospho-Thr-205 and does not recognize unphosphorylated recombinant tau protein. The antibody recognizes multiple closely spaced bands in the region of the blot between 50kDa and 75kDa are detected in adult rat brain lysate, and a ~50kDa band is revealed in embryonic cortical neuron cell culture lysates. Apparently Thr205 is phosphorylated, perhaps constitutively.

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Immunogen: Peptide sequence 190-215 of the human 4 repeat τ with two phosphate groups on Ser 202 and Thr-205, KSGDRSGYSSPGS(PO4)PGT(PO4)PGSRSRTPSL. Peptide included a C-terminal Cys residue to allow coupling to KLH.
HGNC Name: MAPT
UniProt: P10636
Molecular Weight: 48-67 kDa by SDS-PAGE, with higher molecular weight "big tau" isoforms mostly in peripheral tissues.
Host: Mouse
Isotype: IgG1
Species Cross-Reactivity: Human, Rat, Mouse
Format: Protein G affinity purified antibody at 1mg/mL in 50% PBS, 50% glycerol plus 5mM NaN3
Applications: WB, ICC/IF, IHC
Recommended Dilutions: Western blot: 1:10,000. IF/ICC 1:1,000. IHC 1:10,000
Storage: Stable at 4°C for one year, for longer term store at -20°C

      Tau, aka τ, is a low molecular weight member of the microtubule associated protein or MAP family. Several serious human diseases are associated with accumulations of tau protein, most notably the neurofibrillary tangles of Alzheimer's disease. Accumulations of τ in neurons are also characteristic of chronic traumatic encephalopathy, Pick's disease and several other neurodegenerative diseases. Together these disorders are known as "tauopathies". The single mammalian τ gene produces at least 9 different proteins by alternate transcription. In the central nervous system 6 isoforms ranging from 48-67kDa by SDS-PAGE predominate, though larger isotypes are seen mostly in the peripheral nervous system. The τ molecules are very heavily charged and run on SDS-PAGE much more slowly than predicted from their real molecular size. For example the smallest human τ isotype runs at 48kDa on SDS-PAGE but the real molecular weight is 32kDa. Tau proteins are substrates for ser/thr phosphorylation and other post-translational modifications.



Section of rat hippocampuse stained with MCA-1H103 diluted 1:1,000. and detected with DAB (brown) using the Vector Labs ImmPRESS method and reagents with citra buffer retrieval. The antibody staines neuronal perikarya and processes. We have not yet verified that this antibody works on typical human brain sections for IHC.



Dot blots of 2µL of 10mg/mL peptides based on amino acids 190-215 of the human sequence immobiized on nitrocellulose. Peptides were synthesized with with both S202 and T205 phosphorylated (Ser-P202/Thr-P205), just Threonine 205 phosphorylated (Thr-P205), just S202 phosphorylated (Ser-P202) or niether site phosphorylated as shown. Clearly MCA-1H103 strongly recognizes peptides in which T205 is phosphorylated but shows no reactivity with unphosphorylated or Ser202 phosphorylated tau. Binding of MCA-1H103 is also apparently not affected by the state of phosphorylation of Ser202.

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