- Species ReactivityHuman, Mouse, Rat
- SpecificityDetects human, mouse, and rat SHP-2 in Western blots. Does not detect SHP-1 in Western blots.
- SourcePolyclonal Goat IgG
- PurificationAntigen Affinity-purified
- ImmunogenE. coli-derived recombinant human SHP-2
Thr2-Arg593
Accession # NP_002825 - FormulationLyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
- LabelUnconjugated
- Western Blot0.1 µg/mLSee below
- ReconstitutionReconstitute at 0.2 mg/mL in sterile PBS.
- ShippingThe product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
- Stability & StorageUse a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
- Zhao, Z. et al. (1994) J. Biol. Chem. 269:8780.
- Clemmons, D.R. and Maile, L.A. (2005) Mol. Endocrinol. 19:1.
- von Wichert, G. et al. (2003) EMBO J. 22:5023.
- Maroun, C.R. et al. (2000) Mol. Cell. Biol. 20:8513.
- Sugimoto, S. et al. (1993) J. Biol. Chem. 269:22771.
- Long Name:Src Homology Region 2 Domain Phosphatase 2
- Entrez Gene IDs:5781 (Human); 54390 (Mouse); 25622 (Rat)
- Alternate Names:BPTP3; EC 3.1.3.48; MGC14433; protein tyrosine phosphatase, non-receptor type 11; protein tyrosine phosphatase-2; Protein-tyrosine phosphatase 1D; Protein-tyrosine phosphatase 2C; PTP1D; PTP-1D; PTP2C; PTP-2C; PTP2CNS1; PTPN11; SHP2; SHP-2; SHP2CFC; SHPTP2; SH-PTP2; SH-PTP2Noonan syndrome 1; SH-PTP3; tyrosine-protein phosphatase non-receptor type 11
Background:
Src-Homology domain-2 containing protein tyrosine Phosphatase 2 (SHP-2), also called protein tyrosine phosphatase, non-receptor type 11 (PTPN11), PTP1D, PTP2C, and SYP, is an enzyme that dephosphorylates tyrosine residues in proteins. The protein contains two Src homology 2 (SH2) domains, which both regulate the activity of the enzyme (1) and allow it to selectively bind to SH2 sites on proteins such as Dok1, IRS1, and the insulin receptor (2). SHP-2 plays a unique stimulatory role in cell signaling. Cells lacking SHP-2 have poor mobility because the hyper-phosphorylation of FAK and other proteins in the focal adhesion complex (3) prevents turnover of cellular attachment points. Without SHP-2, sustained ERK stimulation does not take place (4). The Y992 phosphorylation site of EGFR is a particularly good substrate for SHP-2 (5) and a phosphopeptide containing this sequence can be used to measure the activity of the enzyme (R&D Systems, Catalog # ES006) by detecting release of phosphate (R&D Systems, Catalog # DY996).
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血清内含有抗体,封闭的时候,选用同物种的血清,比如,使用小鼠单抗的话,就选用小鼠血清来进行封闭。这样,如果细胞表面有可以非特异性结合小鼠抗体的部位都会事先被结合,就不会再结合荧光标记的特异性抗体了。
如果一个抗体可以用来“做western blot,免疫荧光,免疫共沉淀,ELISA”,我估计着是HRP或碱性磷酸酶或生物素之类标记的,然后需要再加荧光标记的二抗,这类抗体也是不能做流式用的。
我只用过流式的抗体来做免疫荧光(直接染色),效果还不错,但是反过来就没试过。
比如,你使用了小鼠IgG1, PE 标记的特异性抗体,那就选用小鼠IgG1, PE标记的非特异性抗体。
下面这个图,是一个比较典型的使用了同型对照的结果图向左转|向右转灰色的曲线是未染色的样本,黄色是同型对照,红色是特异性抗体染色。由于细胞对抗体的非特异性结合,一般同型对照都会比未染色样本的荧光信号要强。所以设置阴性阈值的时候,要根据同型对照,而不是未染色的细胞。
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