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BD Transduction Laboratories™ Purified Mouse Anti-R-PTP-ζ
克隆 12/RPTPb (RUO)
Western blot analysis of R-PTP-ζ in human T leukemia (left). Lysate from Jurkat cells was probed with Purified Mouse Anti-R-PTP-ζ (Cat. No. 610179 or 610180) at dilutions of 1:250, 2:500, and 1:1000 (Lanes 1, 2, and 3, respectively), followed by HRP Goat Anti-Mouse Ig (Cat. No. 610179). R-PTP-ζ is identified as a band of 250 kDa.
Immunofluorescent staining of human neuroblastoma cells (right). SH-SY5Y cells (ATCC CRL-2266) were seeded in a 384-well collagen-coated microplate at ~8,000 cells per well. After overnight incubation, the cells were fixed, permeabilized with cold methanol, and stained with Purified Mouse Anti-R-PTP-ζ. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen, pseudocolored green). Cell nuclei were counterstained with Hoechst 33342 (Cat. No. 561908, pseudocolored blue). The image was taken on a BD Pathway™ 855 or 435 Bioimager System using a 20× objective and merged using BD AttoVision™ software. This antibody also stained SK-N-SH (human neuroblastoma) and C6 (rat glioma) cells using both the Triton X-100 and methanol fix/perm protocols (see Recommended Assay Procedure; Bioimaging protocol link).
Western blot analysis of R-PTP-ζ in human T leukemia (left). Lysate from Jurkat cells was probed with Purified Mouse Anti-R-PTP-ζ (Cat. No. 610179 or 610180) at dilutions of 1:250, 2:500, and 1:1000 (Lanes 1, 2, and 3, respectively), followed by HRP Goat Anti-Mouse Ig (Cat. No. 610179). R-PTP-ζ is identified as a band of 250 kDa.
Immunofluorescent staining of human neuroblastoma cells (right). SH-SY5Y cells (ATCC CRL-2266) were seeded in a 384-well collagen-coated microplate at ~8,000 cells per well. After overnight incubation, the cells were fixed, permeabilized with cold methanol, and stained with Purified Mouse Anti-R-PTP-ζ. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen, pseudocolored green). Cell nuclei were counterstained with Hoechst 33342 (Cat. No. 561908, pseudocolored blue). The image was taken on a BD Pathway™ 855 or 435 Bioimager System using a 20× objective and merged using BD AttoVision™ software. This antibody also stained SK-N-SH (human neuroblastoma) and C6 (rat glioma) cells using both the Triton X-100 and methanol fix/perm protocols (see Recommended Assay Procedure; Bioimaging protocol link).
Western blot analysis of R-PTP-ζ in human T leukemia (left). Lysate from Jurkat cells was probed with Purified Mouse Anti-R-PTP-ζ (Cat. No. 610179 or 610180) at dilutions of 1:250, 2:500, and 1:1000 (Lanes 1, 2, and 3, respectively), followed by HRP Goat Anti-Mouse Ig (Cat. No. 610179). R-PTP-ζ is identified as a band of 250 kDa.
Immunofluorescent staining of human neuroblastoma cells (right). SH-SY5Y cells (ATCC CRL-2266) were seeded in a 384-well collagen-coated microplate at ~8,000 cells per well. After overnight incubation, the cells were fixed, permeabilized with cold methanol, and stained with Purified Mouse Anti-R-PTP-ζ. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen, pseudocolored green). Cell nuclei were counterstained with Hoechst 33342 (Cat. No. 561908, pseudocolored blue). The image was taken on a BD Pathway™ 855 or 435 Bioimager System using a 20× objective and merged using BD AttoVision™ software. This antibody also stained SK-N-SH (human neuroblastoma) and C6 (rat glioma) cells using both the Triton X-100 and methanol fix/perm protocols (see Recommended Assay Procedure; Bioimaging protocol link).
Western blot analysis of R-PTP-ζ in human T leukemia (left). Lysate from Jurkat cells was probed with Purified Mouse Anti-R-PTP-ζ (Cat. No. 610179 or 610180) at dilutions of 1:250, 2:500, and 1:1000 (Lanes 1, 2, and 3, respectively), followed by HRP Goat Anti-Mouse Ig (Cat. No. 610179). R-PTP-ζ is identified as a band of 250 kDa.
Immunofluorescent staining of human neuroblastoma cells (right). SH-SY5Y cells (ATCC CRL-2266) were seeded in a 384-well collagen-coated microplate at ~8,000 cells per well. After overnight incubation, the cells were fixed, permeabilized with cold methanol, and stained with Purified Mouse Anti-R-PTP-ζ. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen, pseudocolored green). Cell nuclei were counterstained with Hoechst 33342 (Cat. No. 561908, pseudocolored blue). The image was taken on a BD Pathway™ 855 or 435 Bioimager System using a 20× objective and merged using BD AttoVision™ software. This antibody also stained SK-N-SH (human neuroblastoma) and C6 (rat glioma) cells using both the Triton X-100 and methanol fix/perm protocols (see Recommended Assay Procedure; Bioimaging protocol link).
Western blot analysis of R-PTP-ζ in human T leukemia (left). Lysate from Jurkat cells was probed with Purified Mouse Anti-R-PTP-ζ (Cat. No. 610179 or 610180) at dilutions of 1:250, 2:500, and 1:1000 (Lanes 1, 2, and 3, respectively), followed by HRP Goat Anti-Mouse Ig (Cat. No. 610179). R-PTP-ζ is identified as a band of 250 kDa.
Immunofluorescent staining of human neuroblastoma cells (right). SH-SY5Y cells (ATCC CRL-2266) were seeded in a 384-well collagen-coated microplate at ~8,000 cells per well. After overnight incubation, the cells were fixed, permeabilized with cold methanol, and stained with Purified Mouse Anti-R-PTP-ζ. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen, pseudocolored green). Cell nuclei were counterstained with Hoechst 33342 (Cat. No. 561908, pseudocolored blue). The image was taken on a BD Pathway™ 855 or 435 Bioimager System using a 20× objective and merged using BD AttoVision™ software. This antibody also stained SK-N-SH (human neuroblastoma) and C6 (rat glioma) cells using both the Triton X-100 and methanol fix/perm protocols (see Recommended Assay Procedure; Bioimaging protocol link).
Western blot analysis of R-PTP-ζ in human T leukemia (left). Lysate from Jurkat cells was probed with Purified Mouse Anti-R-PTP-ζ (Cat. No. 610179 or 610180) at dilutions of 1:250, 2:500, and 1:1000 (Lanes 1, 2, and 3, respectively), followed by HRP Goat Anti-Mouse Ig (Cat. No. 610179). R-PTP-ζ is identified as a band of 250 kDa.
Immunofluorescent staining of human neuroblastoma cells (right). SH-SY5Y cells (ATCC CRL-2266) were seeded in a 384-well collagen-coated microplate at ~8,000 cells per well. After overnight incubation, the cells were fixed, permeabilized with cold methanol, and stained with Purified Mouse Anti-R-PTP-ζ. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen, pseudocolored green). Cell nuclei were counterstained with Hoechst 33342 (Cat. No. 561908, pseudocolored blue). The image was taken on a BD Pathway™ 855 or 435 Bioimager System using a 20× objective and merged using BD AttoVision™ software. This antibody also stained SK-N-SH (human neuroblastoma) and C6 (rat glioma) cells using both the Triton X-100 and methanol fix/perm protocols (see Recommended Assay Procedure; Bioimaging protocol link).
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商品通知
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Triton is a trademark of the Dow Chemical Company.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
配套商品
Receptor-type tyrosine-protein phosphatase zeta (PTPRZ, R-PTP-ζ, or PTPζ) was previously known as RPTPβ or RPTP ζ /β. It is the first mammalian tyrosine phosphatase to be characterized whose expression is limited to the nervous system. R-PTP-ζ consists of a large extracellular domain, a single transmembrane domain, and a cytoplasmic portion with two tandem catalytic domains. Three forms of R-PTP-ζ exist which appear to be derived from alternative splicing. The 9.5-kb and 6.4-kb transcripts encode two transmembrane forms. The 8.5 kb transcript encodes a secreted form of the extracellular domain of R-PTP-ζ. A region of 266 amino acids in the extracellular domain shows a high degree of homology with carbonic anhydrase. This region is very similar to rat brain chondroitin sulfate proteoglycan (3F8 PG) which appears to be the rat homologue of the entire extracellular domain of human R-PTP-ζ. The apparent molecular weight of human R-PTP-ζ is 250 kDa and approaches 300 kDa when the protein is glycosylated. R-PTP-ζ has regulatory effects in the development and repair of the nervous system. Interactions of R-PTP-ζ with the cytokines MK and PTN regulate cellular adhesion, motility, and migration events in nervous system development, and dis-regulation of these interactions appear to be involved in some neurologic disorders.
The 12/RPTPb monoclonal antibody was generated against a region known to have a high degree of homology to other proteoglycans, such as R-PTP-γ (75% homology). The immunogen sequence is not present in R-PTP-β, also known as VE-PTP or PTPRB, so cross-reactivity is not expected.
研发参考 (13)
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Fukazawa N, Yokoyama S, Eiraku M, Kengaku M, Maeda N. Receptor type protein tyrosine phosphatase zeta-pleiotrophin signaling controls endocytic trafficking of DNER that regulates neuritogenesis.. Mol Cell Biol. 2008; 28(14):4494-506. (Clone-specific). 查看参考
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Hayashi N, Oohira A, Miyata S. Synaptic localization of receptor-type protein tyrosine phosphatase zeta/beta in the cerebral and hippocampal neurons of adult rats.. Brain Res. 2005; 1050(1-2):163-9. (Clone-specific). 查看参考
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Herradón G, Ezquerra L. Blocking receptor protein tyrosine phosphatase beta/zeta: a potential therapeutic strategy for Parkinson's disease.. Curr Med Chem. 2009; 16(25):3322-9. (Biology). 查看参考
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Herradón G, Pérez-García C. Targeting midkine and pleiotrophin signalling pathways in addiction and neurodegenerative disorders: recent progress and perspectives.. Br J Pharmacol. 2014; 171(4):837-48. (Biology). 查看参考
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Kowalik L, Hudspeth AJ. A search for factors specifying tonotopy implicates DNER in hair-cell development in the chick's cochlea.. Dev Biol. 2011; 354(2):221-31. (Clone-specific). 查看参考
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Krueger NX, Saito H. A human transmembrane protein-tyrosine-phosphatase, PTP zeta, is expressed in brain and has an N-terminal receptor domain homologous to carbonic anhydrases. Proc Natl Acad Sci U S A. 1992; 89(16):7417-7421. (Biology). 查看参考
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Levy JB, Canoll PD, Silvennoinen O, et al. The cloning of a receptor-type protein tyrosine phosphatase expressed in the central nervous system. J Biol Chem. 1993; 268(14):10573-10581. (Biology). 查看参考
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Maeda N, Noda M. Involvement of receptor-like protein tyrosine phosphatase zeta/RPTPbeta and its ligand pleiotrophin/heparin-binding growth-associated molecule (HB-GAM) in neuronal migration. J Cell Biol. 1998; 142(1):203-216. (Biology: Immunofluorescence). 查看参考
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Meng K, Rodriguez-Peña A, Dimitrov T, et al. Pleiotrophin signals increased tyrosine phosphorylation of beta beta-catenin through inactivation of the intrinsic catalytic activity of the receptor-type protein tyrosine phosphatase beta/zeta.. Proc Natl Acad Sci U S A. 2000; 97(6):2603-8. (Clone-specific). 查看参考
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Padilla PI, Wada A, Yahiro K, et al. Morphologic differentiation of HL-60 cells is associated with appearance of RPTPbeta and induction of Helicobacter pylori VacA sensitivity. J Biol Chem. 2000; 275(20):15200-15206. (Clone-specific). 查看参考
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Ratcliffe CF, Qu Y, McCormick KA, et al. A sodium channel signaling complex: modulation by associated receptor protein tyrosine phosphatase beta. Nat Neurosci. 2000; 3(5):437-444. (Clone-specific). 查看参考
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Shi Y, Ping YF, Zhou W, et al. Tumour-associated macrophages secrete pleiotrophin to promote PTPRZ1 signalling in glioblastoma stem cells for tumour growth.. Nat Commun. 2017; 8:15080. (Clone-specific). 查看参考
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den Hertog J, Blanchetot C, Buist A, Overvoorde J, van der Sar A, Tertoolen LG. Receptor protein-tyrosine phosphatase signalling in development.. Int J Dev Biol. 1999; 43(7):723-33. (Biology). 查看参考
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