Recombinant Human His6-UAF1 Protein, CF Summary
Recombinant Human UAF1 is an activator of Ubiquitin-specific deconjugating enzymes USP1, USP12, and USP46. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human UAF1 concentration equal to the concentration of deconjugating enzyme present(1:1 stoichiometry) in in vitro deconjugation reactions utilizing Ubiquitin-AMC.
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins.Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration.The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard.In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
E-566
| Formulation | X mg/ml (X μM) in 50 mM HEPES, pH 7.5, 100 mM NaCl, 10% glycerol, 1 mM TCEP |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Reconstitution Calculator
Background: UAF1
UAF1 (USP1-Associated Factor 1, also known as WD Repeat-containing protein 48 or WDR48) is a positive regulator of at least three deubiquitinating enzymes of the C19 peptidase family including USP1, USP12, and USP46. In the absence of UAF1, all three of these deubiquitinases are almost completely inactive, and available biochemical evidence suggests UAF1 activation occurs by increasing catalytic turnover as opposed to altered substrate affinity (at least in the case of UAF1/USP1) of the enzymes. Most notably, UAF1 strongly activates USP1 deubiquitination of monoubiquitinated FANCD2 and PCNA, thereby playing important roles in DNA damage response and translesion synthesis. The UAF1/USP12 complex has recently been described as a negative regulator of Notch signaling. Disruption of UAF1-dependent USP12 activity interrupts Notch trafficking to the lysosomes, leading to an increased amount of receptor at the cell surface and to higher Notch activity. At the biochemical level, UAF1/USP12 deubiquitinates the non-activated form of Notch in cell culture and in vitro. This recombinant protein contains a C-terminal 6-His tag.
- Cohn M.A., et al. (2009) J Biol. Chem. 284: 5343-5351
- Huang T.T., et al. (2006) Nat. Cell Biol. 8: 339-347
- Joo H-Y., et al. (2011) Biol. Chem. 286: 7190-7201
- Moretti J., et al. (2012) J Biol. Chem. 287: 29429-29441
- Williams S.A, et al. (2011) Cell 146: 918–930
Citation for Recombinant Human His6-UAF1 Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products.The data collected includes not only links to publications in PubMed,but also provides information about sample types, species, and experimental conditions.
1Citation: Showing 1 - 1
- Site-specific ubiquitylation and SUMOylation using genetic-code expansion and sortaseAuthors: M Fottner, AD Brunner, V Bittl, D Horn-Ghetk, A Jussupow, VRI Kaila, A Bremm, K LangNat. Chem. Biol., 2019;15(3):276-284.Species: HumanSample Types: Recombinant ProteinApplications: Ubiquitination
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多谢！
不知道能不能直接买到大鼠抗豚鼠血管IgG的ELISA试剂盒。
如果买不到这种试剂盒，用空白板的话（就是反应孔中没有抗原或者抗体包被），把自制的豚鼠血管抗原加到反应孔中进行孵育，能不能使抗原附着在孔壁上？这样的话就可以按照elisa的操作步骤进行大鼠血清检测了。
目前的设想是自制抗原，粉碎豚鼠的血管，制成悬液，通过反复多次的冻融，离心后收集上清液作为抗原（即豚鼠血管抗原）。
然后将抗原加入到elisa的反应孔中（这种是特制的空白板，就是反应孔中没有抗原或抗体包被）进行孵育，使豚鼠血管抗原附着在孔壁上，就是让反应孔充当固相载体，形成固相抗原。倒掉多余的抗原。
再加入待检测的大鼠血清，这样血清中的特异性抗豚鼠血管IgG就可以跟固相的抗原结合，形成固相抗原抗体复合物。
加入酶标的兔抗鼠或者羊抗鼠IgG,形成酶标的抗原抗体复合物。
然后就是一些显色步骤。
现在关键的问题就是抗原加入空白板，能不能形成固相抗原，如果不能跟孔壁附着的话，在后面洗涤的过程中就被洗掉了，那就没办法完成检测了。
请各位大侠给指条明路啊！！！小弟在此多谢了！！！
