STEP Antibody (23E5)

Western Blot: STEP Antibody (23E5) [NB300-202] - Analysis of STEP protein in striatal rat protein homogenates

Immunohistochemistry-Paraffin: STEP Antibody (23E5) [NB300-202] - STEP was detected in immersion fixed paraffin-embedded sections of human brain basal ganglia using Mouse Anti-Human STEP (23E5) Monoclonal Antibody (Catalog # NB300-202) at 1:100 for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm in neurons.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ICC/IF, IHC, IHC-Fr, IHC-P, IP
• Clone: 23E5
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

STEP Antibody (23E5) Summary

• Immunogen: A synthetic peptide of rat STEP46 (amino acids 57-74: MCTPGCNEEGFGYLVSPR)
• Localization: Cytoplasmic
• Specificity: NB300-202 is specific for STEP61, STEP46, STEP38, and STEP20.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: PTPN5
• Purity: Unpurified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:1000 -1:2000
  • Immunocytochemistry/Immunofluorescence 1:100-1:500
  • Immunohistochemistry 1:100-1:500
  • Immunohistochemistry-Frozen 1:100-1:500
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation 1:50
• Application Notes: This STEP (23E5) antibody is useful for Western blot, Immunoprecipitation, Immunocytochemistry/Immunofluorescence and Immunohistochemistry on frozen sections. By Western Blot, this STEP antibody recognizes bands at ~64-66, 46, 37, and 33 kDa, in rat brain tissues. Neuronal cells have been used for ICC/IF and frozen brain sections have been used for IHC.
• Control:

  Rat Brain Whole Tissue Lysate (Adult Whole Normal)

Reactivity Notes

Human, mouse, rat.

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: Ascites
• Preservative: 0.09% Sodium Azide
• Purity: Unpurified

Alternate Names for STEP Antibody (23E5)

• EC 3.1.3.48
• FLJ14427
• Neural-specific protein-tyrosine phosphatase
• protein tyrosine phosphatase, non-receptor type 5 (striatum-enriched)
• protein-tyrosine phosphatase striatum-enriched
• STEPPTPSTEP
• Striatum-enriched protein-tyrosine phosphatase
• tyrosine-protein phosphatase non-receptor type 5

Background

STEP (STriatal-Enriched protein tyrosine Phosphatase; PTPN5) is a brain-specific phosphatase enzyme which is regarded as an "LTD protein" because it mediates AMPA receptor internalization during metabotropic glutamate receptor-dependent long-term depression (mGluR-LTD). STEP regulates MAPKs, Src kinases and NMDA receptors which are involved in synaptic plasticity as well as in neuronal cell survival mechanisms. STEP is highly expressed in striatum and is also found in hippocampus, cortex, amygdala, optic nerve, and spinal cord, but not expressed in cerebellum. PKA mediated phosphorylation at Ser-245 site deactivates STEP whereas MAPKs regulated phosphorylation at Thr-255/Ser-268 stabilizes STEP through preventing its proteasomal degradation. Out of four alternatively spliced variants (STEP61, STEP46, STEP38, STEP20), two major isoforms namely STEP61 and STEP46, contains a signature consensus tyrosine phosphatase sequence, [I/V]HCxAGxxR[S/T]G, necessary for its catalytic activity, and a kinase-interacting motif (KIM) essential for substrate binding. During extrasynaptic NMDAR stimulation, a truncated STEP product, STEP33, is generated by calpain-mediated cleavage between residues Ser224 and Leu225 in the KIM domain leading to disruption of STEP's ability to associate with and dephosphorylate its substrates. Subsequently, proteolytic cleavage of STEP after extrasynaptic NMDAR stimulation results in p38 activation, and initiates cell death signaling cascade. STEP participates in dopaminergic, glutamatergic, and nicotinic signaling, thereby playing a key role in psychostimulant addiction and Alzheimer's disease.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for STEP Antibody (NB300-202)(32)

Publications using NB300-202

• Evans C, Thomas R, Freeman T et al. Selective reduction of APP-BACE1 activity improves memory via NMDA NR2B receptor-mediated mechanisms in aged PDAPP mice. Int J Mol Sci. Nov 5 2018
• Carvajal FJ, Mira RG, Rovegno M et al. Age-related NMDA signaling alterations in SOD2 deficient mice. Biochim. Biophys. Acta. Mar 22 2018 [PMID: 29577983] (ICC/IF)
• Pose-Utrilla J, Garcia-Guerra L, Del Puerto A et al. Excitotoxic inactivation of constitutive oxidative stress detoxification pathway in neurons can be rescued by PKD1. Nat Commun 2017 Dec 22 [PMID: 29273751] (Mouse)
• Won S, Incontro S, Nicoll RA, Roche KW. PSD-95 stabilizes NMDA receptors by inducing the degradation of STEP61 Proc. Natl. Acad. Sci. U.S.A. 2016 Jul 25 [PMID: 27457929] (WB)
• Rajagopal S, Deb I, Poddar R, Paul S. Aging is associated with dimerization and inactivation of the brain-enriched tyrosine phosphatase STEP. Neurobiology of Aging. 2016 Feb 12 [PMID: 27103516] (IP, Rat)
• Poddar R, Rajagopal S, Shuttleworth CW, Paul S. Zn2+-dependent activation of the Trk signaling pathway induces phosphorylation of the brain-enriched tyrosine phosphatase STEP: molecular basis for Zn2+-induced ERK MAPK activation. J. Biol. Chem. 2015 Nov 16 [PMID: 26574547] (WB, IP, Rat)
• Kim HN, Jang JY, Choi BT. A single fraction from Uncaria sinensis exerts neuroprotective effects against glutamate-induced neurotoxicity in primary cultured cortical neurons Anat Cell Biol 2015 Jun 01 [PMID: 26140220] (WB, Rat)
  
  Details:
  STEP antibody was used for WB assays on rat's primary cortical neurons which were treated with hexane extract from Uncaria sinensis (JGH43IA 0.1 ug/ml) for 24 hours followed by 300uM of glutamate for 15 minutes (Fig. 5 ).
• Wang Y, Briz V, Chishti A et al. Distinct Roles for u-Calpain and m-Calpain in Synaptic NMDAR-Mediated Neuroprotection and Extrasynaptic NMDAR-Mediated Neurodegeneration. J Neurosci. 2013 Nov 27 [PMID: 24285894] (WB, Rat)
• Deb I, Manhas N, Poddar R et al. Neuroprotective Role of a Brain-Enriched Tyrosine Phosphatase, STEP, in Focal Cerebral Ischemia. J Neurosci. 2013 Nov 6 [PMID: 24198371] (IP, WB, Rat)
• Kim HN, Kim YR, Jang JY et al. Neuroprotective effects of Polygonum multiflorum extract against glutamate-induced oxidative toxicity in HT22 hippocampal cells. J Ethnopharmacol. 2013 Aug 22 [PMID: 23973786] (WB, Mouse)
• Sun WL, Zelek-Molik A, McGinty JFet al. Short and long access to cocaine self-administration activates tyrosine phosphatase STEP and attenuates GluN expression but differentially regulates GluA expression in the prefrontal cortex. Psychopharmacology (Berl) 2013 Apr 27 [PMID: 23624776] (IP, Rat)
• Shinohara Y, Hosoya A, Yahagi K, Ferecsko AS, Yaguchi K, Sik A, Itakura M, Takahashi M, Hirase H. Hippocampal CA3 and CA2 have distinct bilateral innervation patterns to CA1 in rodents. Eur J Neurosci;35(5):702-710. doi: 10.1111/j.1460-9568.2012.07993.x. 2012 Mar. [PMID: 22339771]
• Farooq M, Kaswala RH, Kleiman NJ, Kasinathan C, Frederikse PH. GluA2 AMPA glutamate receptor subunit exhibits codon 607 Q/R RNA editing in the lens. Biochem Biophys Res Commun;418(2):273-7. 2012 Feb 10. [PMID: 22266371]
• Davis MM, Olausson P, Greengard P, Taylor JR, Nairn AC. Regulator of calmodulin signaling knockout mice display anxiety-like behavior and motivational deficits. Eur J Neurosci;35(2):300-8. 2012 Jan. [PMID: 22250817] (WB, Mouse)
• Mukherjee S, Poddar R, Deb I, Paul S. Dephosphorylation of specific sites in the KIS domain leads to ubiquitin-mediated degradation of the tyrosine phosphatase STEP. Biochem J. 2011 Jul 21. [PMID: 21777200]
• Poddar R, Deb I, Mukherjee S, Paul S. NR2B-NMDA receptor mediated modulation of the tyrosine phosphatase STEP regulates glutamate induced neuronal cell death. J Neurochem 2010 Dec [PMID: 21029094] (WB, Rat)
• Zhang Y, Kurup P, Xu J et al. Genetic reduction of striatal-enriched tyrosine phosphatase (STEP) reverses cognitive and cellular deficits in an Alzheimer's disease mouse model. Proc Natl Acad Sci U S A;107(44):19014-9. 2010 Nov 2. [PMID: 20956308] (WB, IP, Mouse)
• Kurup P, Zhang Y, Xu J, Venkitaramani DV, Haroutunian V, Greengard P, Nairn AC, Lombroso PJ. Abeta-mediated NMDA receptor endocytosis in Alzheimer's disease involves ubiquitination of the tyrosine phosphatase STEP61. J Neurosci;30(17):5948-57. 2010 Apr 28. [PMID: 20427654] (WB)
• DiRocco DP, Scheiner ZS, Sindreu CB et al. A Role for Calmodulin-Stimulated Adenylyl Cyclases in Cocaine Sensitization. J Neurosci;29(8):2393-403. 2009 Feb 25. [PMID: 19244515] (ICC/IF, Mouse)
• Goebel-Goody SM, Davies KD, Alvestad Linger RM, Freund RK, Browning MD. Phospho-regulation of synaptic and extrasynaptic N-methyl-d-aspartate receptors in adult hippocampal slices. Neuroscience;158(4):1446-59. 2009 Feb 18. [PMID: 19041929]
• Zhang Y, Venkitaramani DV, Gladding CM, Zhang Y, Kurup P, Molnar E, Collingridge GL, Lombroso PJ. The tyrosine phosphatase STEP mediates AMPA receptor endocytosis after metabotropic glutamate receptor stimulation. J Neurosci;28(42):10561-6. 2008 Oct 15. [PMID: 18923032] (WB, IHC, Rat, Mouse)
• Zhao, M et al. Synaptic plasticity (and the lack thereof) in hippocampal CA2 neurons. J Neurosci;27(44):12025-32. 2007 Oct 31. [PMID: 17978044]
• Hu Y, Zhang Y, Venkitaramani DV, Lombroso PJ. Translation of striatal-enriched protein tyrosine phosphatase (STEP) after beta1-adrenergic receptor stimulation. J Neurochem;103(2):531-41. 2007 Oct. [PMID: 17623046] (WB, IHC-Fr, Rat)
• Choi, YS et al. Status epilepticus-induced somatostatinergic hilar interneuron degeneration is regulated by striatal enriched protein tyrosine phosphatase. J Neurosci;27(11):2999-3009. 2007 Mar 14. [PMID: 17360923] (IHC, Mouse)
• Paul S, Nairn AC, Wang P, Lombroso PJ. NMDA-mediated activation of the tyrosine phosphatase STEP regulates the duration of ERK signaling. Nat Neurosci;6(1):34-42. 2003 Jan. [PMID: 12483215] (WB, IP)
• Nguyen TH, Liu J, Lombroso PJ. Striatal enriched phosphatase 61 dephosphorylates Fyn at phosphotyrosine 420. J Biol Chem;277(27):24274-9. 2002 Jul 5. [PMID: 11983687] (WB)
• Pelkey KA, Askalan R, Paul S, Kalia LV, Nguyen TH, Pitcher GM, Salter MW, Lombroso. Tyrosine phosphatase STEP is a tonic brake on induction of long-term potentiation. Neuron. 34(1):127-38. 2002 Mar 28. [PMID: 11931747]
• Paul S, Snyder GL, Yokakura H, Picciotto MR, Nairn AC, Lombroso PJ. The Dopamine/D1 receptor mediates the phosphorylation and inactivation of the protein tyrosine phosphatase STEP via a PKA-dependent pathway. J Neurosci;20(15):5630-8. 2000 Aug 1. [PMID: 10908600] (WB, IP, Rat )
• Nguyen TH, Paul S, Xu Y, Gurd JW, Lombroso PJ. Calcium-dependent cleavage of striatal enriched tyrosine phosphatase (STEP). J Neurochem;73(5):1995-2001. 1999 Nov. [PMID: 10537058]
• Gurd JW, Bissoon N, Nguyen TH, Lombroso PJ, Rider CC, Beesley PW, Vannucci SJ. Hypoxia-ischemia in perinatal rat brain induces the formation of a low molecular weight isoform of striatal enriched tyrosine phosphatase (STEP). J Neurochem;73(5):1990-4. 1999 Nov. [PMID: 10537057]
• Bult A, Zhao F, Dirkx R Jr, Sharma E, Lukacsi E, Solimena M, Naegele JR, Lombroso PJ. STEP61: a member of a family of brain-enriched PTPs is localized to the endoplasmic reticulum. J Neurosci;16(24):7821-31. 1996 Dec 15. [PMID: 8987810] (IHC-Fr, WB, Rat)
• Boulanger LM, Lombroso PJ, Raghunathan A, During MJ, Wahle P, Naegele JR. Cellular and molecular characterization of a brain-enriched protein tyrosine phosphatase. J Neurosci;15(2):1532-44. 1995 Feb. [PMID: 7869116]

Review for STEP Antibody (NB300-202) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Anonymous WB Mouse 12/23/2014

Summary

• Application: Western Blot
• Sample Tested: See PMID 22266371
• Species: Mouse

Blocking

• Blocking Details: See PMID 22266371

Details

• Detection Notes: See PMID 22266371

Comments

• Comments: Independent review from antibodyresource.comPublished in PMID 22266371.

FAQs for STEP Antibody (NB300-202). (Showing 1 - 1 of 1 FAQs).

1. Is it possible this antibody can detect phospho form of that? Some paper shows that phosphorylated STEP was detected by an pware shift in the mobility of the STEP band. The paper is 17814, The Journal of Neuroscience, November 6, 2013, 33(45):17814 -17826.
   • This antibody may detect both phosphorylated and non-phosphorylated STEP although it has never specifically been tested for this.

Bioinformatics

• Gene Symbol: PTPN5
• Entrez:
  • 84867(Human)
  • 29644(Rat)