AQUApure Linear Tetra-Ub Chains (M1-link) Protein, CF Summary
Ubiquitin chains vary in length, linkage, and function. Linear (Met1)-linked Tetra-Ubiquitin Chains (Ub4) are ideal for investigating Ubiquitin-binding proteins and as substrates for Ubiquitin-specific isopeptidases. Reaction conditions will need to be optimized for each specific application. IMPORTANT: Heating this product in SDS-PAGE buffer or terminating reactions containing this product with heated SDS-PAGE buffer could lead to unexpected, high apparent molecular weight banding or smearing on gels that is not representative of product purity. For optimal results, we recommend incubation in SDS-PAGE buffer + DTT at <40 °c="" for="" 20="" minutes="" prior="" to="" gel="">
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.
UC-710B
| Formulation | 1 mg/ml (29 μM) in sterile, deionized water |
| Shipping | The product is shipped with polar packs. 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: Tetra-Ubiquitin
Linkage specific Poly-Ubiquitin chains may be used as a substrate for in vitro reactions with deubiquitinating enzymes ("DUB"s") that cleave the peptide or isopeptide linkage between adjacent Ubiquitin molecules. Poly-Ubiquitin chains can also be used to investigate mechanisms of binding and recognition between the chains and other proteins that contain Ubiquitin-Associated domains (UBAs), Ubiquitin-interacting motifs (UIMs), ZnF"s and/or other Ubiquitin-sensing elements.
Linear ("M1")-linked Tetra-Ubiquitin chains are manufactured using recombinant methods to avoid the potential for contaminating synthetic intermediates. The correctness of linkage and purity of each production lot is assessed using the Absolute Quantitation of Ubiquitin method (Ub-AQUA), an LCMS-based technique that provides extremely accurate information on the composition of Poly-Ubiquitin samples.
- Kirkpatrick D.S., et al. (2006) Nat Cell Biol. 8(7): 700-10
- Ordureau, A., et al. (2014) Mol. Cell 56(3): 360–375
- Ordureau, A., et al. (2015) Pro. Nat. Acad. of Sci. USA 112(21): 6637–6642
- Phu L., et al. (2011) Mol Cell Proteomics 10(5): M110.003756
Citations for AQUApure Linear Tetra-Ub Chains (M1-link) 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.
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- ZUFSP Deubiquitylates K63-Linked Polyubiquitin Chains to Promote Genome StabilityAuthors: P Haahr, N Borgermann, X Guo, D Typas, D Achuthanku, S Hoffmann, R Shearer, TK Sixma, N MailandMol. Cell, 2018;0(0):.Species: HumanSample Types: Cell LysatesApplications: Bioassay
- CP5 system, for simple and highly efficient protein purification with a C-terminal designed mini tagAuthors: H Takeda, W Zhou, K Kido, R Suno, T Iwasaki, T Kobayashi, T SawasakiPLoS ONE, 2017;12(5):e0178246.Species: N/ASample Types: ProteinApplications: Enzyme Assay
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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！！！
