Anti-ErbB2 Affibody®分子(Agarose) (ab31892)

概述

  • 产品名称Anti-ErbB2 Affibody®分子(Agarose)
    参阅全部 ErbB 2 affibody®分子
  • 偶联物Agarose
  • 共轭说明This molecule is immobilized on agarose at the unique C-terminal cysteine.
  • 特异性This product binds to the extracellular domain of human ErbB2. This agarose immobilized Anti-ErbB2 Affibody® molecule is excellent for immunoprecipitation studies of ErbB2 in cell extracts or other solutions that contain ErbB2 proteins. In addition, the Anti-ErbB2 Affibody® molecule may also be used for affinity chromatography.
  • 经测试应用适用于: IPmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    -

  • 常规说明This product is a recombinant protein produced in E.coli.


    What are Affibody Molecules?
    Affibody® affinity ligands are unique research reagents, produced using innovative protein-engineering technologies. They are small, simple proteins composed of a three-helix bundle based on the scaffold of one of the IgG-binding domains of Protein A. Protein A is a surface protein from the bacterium Staphylococcus aureus. This scaffold has excellent features as an affinity ligand and can be designed to bind with high affinity to any given target protein. The domain consists of 58 amino acids, 13 of which are randomized to generate Affibody® libraries with a large number of ligand variants. Thus, the libraries consist of a multitude of protein ligands with an identical backbone and variable surface-binding properties. In function, Affibody® Molecules mimic monoclonal antibodies. Compared to antibodies, the most striking dissimilarity of Affibody® Molecules is the small size. Affibody® Molecules have a molecular weight of 14kDa, compared to the molecular weight of antibodies, which is 150kDa. In spite of its small size, the binding site of Affibody® Molecules is similar to that of an antibody. The advantages of Affibody® Molcules over antibodies are: -their small size -the simple structure of the molecules -its robust physical properties; able to withstand a broad range of analytical conditions, including extreme pH and elevated temperature -its ability to fold correctly intracellularly -the fast and cost effective production in bacteria -the potential to couple Affibody® Molecules in multimeric constructs Affibody® Molecules have highly competitive properties for applications within affinity purification, sample preparation, protein detection and in vitro diagnostics.

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • 存储溶液Preservative: 0.02% Sodium Azide
    Constituents: 50% slurry in 20mM Sodium phosphate, 150mM Sodium chloride, pH 7.5
  • Concentration information loading...
  • 纯化说明The purity of this product is >98% as determined by SDS-PAGE and RP-HPLC analysis.
  • Affibody®分子说明What are Affibody Molecules?
    Affibody® affinity ligands are unique research reagents, produced using innovative protein-engineering technologies. They are small, simple proteins composed of a three-helix bundle based on the scaffold of one of the IgG-binding domains of Protein A. Protein A is a surface protein from the bacterium Staphylococcus aureus. This scaffold has excellent features as an affinity ligand and can be designed to bind with high affinity to any given target protein. The domain consists of 58 amino acids, 13 of which are randomized to generate Affibody® libraries with a large number of ligand variants. Thus, the libraries consist of a multitude of protein ligands with an identical backbone and variable surface-binding properties. In function, Affibody® Molecules mimic monoclonal antibodies. Compared to antibodies, the most striking dissimilarity of Affibody® Molecules is the small size. Affibody® Molecules have a molecular weight of 14kDa, compared to the molecular weight of antibodies, which is 150kDa. In spite of its small size, the binding site of Affibody® Molecules is similar to that of an antibody. The advantages of Affibody® Molcules over antibodies are: -their small size -the simple structure of the molecules -its robust physical properties; able to withstand a broad range of analytical conditions, including extreme pH and elevated temperature -its ability to fold correctly intracellularly -the fast and cost effective production in bacteria -the potential to couple Affibody® Molecules in multimeric constructs Affibody® Molecules have highly competitive properties for applications within affinity purification, sample preparation, protein detection and in vitro diagnostics.
  • 研究领域
  • 功能Protein tyrosine kinase that is part of several cell surface receptor complexes, but that apparently needs a coreceptor for ligand binding. Essential component of a neuregulin-receptor complex, although neuregulins do not interact with it alone. GP30 is a potential ligand for this receptor. Regulates outgrowth and stabilization of peripheral microtubules (MTs). Upon ERBB2 activation, the MEMO1-RHOA-DIAPH1 signaling pathway elicits the phosphorylation and thus the inhibition of GSK3B at cell membrane. This prevents the phosphorylation of APC and CLASP2, allowing its association with the cell membrane. In turn, membrane-bound APC allows the localization of MACF1 to the cell membrane, which is required for microtubule capture and stabilization.
    In the nucleus is involved in transcriptional regulation. Associates with the 5'-TCAAATTC-3' sequence in the PTGS2/COX-2 promoter and activates its transcription. Implicated in transcriptional activation of CDKN1A; the function involves STAT3 and SRC. Involved in the transcription of rRNA genes by RNA Pol I and enhances protein synthesis and cell growth.
  • 组织特异性Expressed in a variety of tumor tissues including primary breast tumors and tumors from small bowel, esophagus, kidney and mouth.
  • 疾病相关Hereditary diffuse gastric cancer
    Glioma
    Ovarian cancer
    Lung cancer
    Gastric cancer
    Chromosomal aberrations involving ERBB2 may be a cause gastric cancer. Deletions within 17q12 region producing fusion transcripts with CDK12, leading to CDK12-ERBB2 fusion leading to truncated CDK12 protein not in-frame with ERBB2.
  • 序列相似性Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
    Contains 1 protein kinase domain.
  • 翻译后修饰Autophosphorylated. Autophosphorylation occurs in trans, i.e. one subunit of the dimeric receptor phosphorylates tyrosine residues on the other subunit (Probable). Ligand-binding increases phosphorylation on tyrosine residues (PubMed:27134172). Signaling via SEMA4C promotes phosphorylation at Tyr-1248 (PubMed:17554007). Dephosphorylated by PTPN12 (PubMed:27134172).
  • 细胞定位Cytoplasm. Nucleus and Cell membrane. Cytoplasm, perinuclear region. Nucleus. Translocation to the nucleus requires endocytosis, probably endosomal sorting and is mediated by importin beta-1/KPNB1.
  • Information by UniProt
  • 别名
    • Verb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog
    • C erb B2/neu protein
    • CD340
    • CD340 antigen
    • Cerb B2/neu protein
    • CerbB2
    • Erb b2 receptor tyrosine kinase 2
    • ERBB2
    • ERBB2_HUMAN
    • HER 2
    • HER 2/NEU
    • HER2
    • Herstatin
    • Human epidermal growth factor receptor 2
    • Metastatic lymph node gene 19 protein
    • MLN 19
    • MLN19
    • NEU
    • NEU proto oncogene
    • Neuro/glioblastoma derived oncogene homolog
    • Neuroblastoma/glioblastoma derived oncogene homolog
    • NGL
    • p185erbB2
    • Proto-oncogene c-ErbB-2
    • Proto-oncogene Neu
    • Receptor tyrosine-protein kinase erbB-2
    • TKR1
    • Tyrosine kinase type cell surface receptor HER2
    • Tyrosine kinase-type cell surface receptor HER2
    • V erb b2 avian erythroblastic leukemia viral oncogene homolog 2
    • V erb b2 avian erythroblastic leukemia viral oncogene homolog 2 (neuro/glioblastoma derived oncogene homolog)
    • V erb b2 avian erythroblastic leukemia viral oncoprotein 2
    • V erb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog
    • V erb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog (avian)
    • Verb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog (avian)
    see all
  • 数据库链接

应用

Our Abpromise guarantee covers the use of ab31892 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IP Use at an assay dependent dilution.
AFFIBODY® MOLECULES ARE PROTOCOL SPECIFIC. PLEASE REFER TO THE "PROTOCOLS" SECTION.

Anti-ErbB2 Affibody® Molecule (Agarose) 图像

  • The agarose immobilized Anti-ErbB2 Affibody® molecule precipitates ErbB2-protein from cell extracts derived from ErbB2 positive cell lines SK-BR-3 and HEP-G2, but not from the ErbB2 low expressing SH-SY5Y cell line.

    RESULTS

    IMMUNOPRECIPITATION

    Cell extracts prepared from SK-BR3, HEP-G2 and SH-SY5Y cells were incubated with agarose immobilized Anti-ErbB2 Affibody® molecule for 2 hours. After incubation, the unbound proteins were washed away and the bound protein was eluted with SDS-PAGE separation and blotted onto a PVDF filter. The filter was stained with an antibody against the intracellular domain of the ErbB2 receptor. The antibody binds to both the fulllength protein (150 kDa) and to a splice variant (approximately 100 kDa) of the receptor that has been reported to be produced by several cell lines.

    Figure 1a shows that the ErbB2 proteins were precipitated from different volumes of SK-BR3 cell extract using the agarose immobilized Anti-ErbB2 Affibody® molecule. An intense ErbB2 band was obtained on the Western blot from as little as 25 μl extract. With increased volume of extract, the ErbB2 band became even more intense. ErbB2 proteins were also precipitated from HEP-G2 cell extract, show in figure 1b. A relatively strong band was obtained when precipitating from 300 μl extract indicating that HEP-G2 expresses ErbB2 to a lower level than SK-BR3. No bands were visible after precipitation with SH-SY5Y cell extracts, shown in figure 1 c, suggesting that this cell line has a very low level of ErbB2 expression.

    These results show that the Anti-ErbB2 Affibody® molecule efficiently precipitates ErbB2 proteins from a complex mix of proteins. When performing immunoprecipitation experiments with antibodies, there is often a problem with cross reactions between the enzyme conjugated second step reagent and the precipitating antibody. This type of cross reaction is avoided with an Affibody® molecule as the precipitating reagent.

Anti-ErbB2 Affibody® Molecule (Agarose) (ab31892)参考文献

This product has been referenced in:
  • Pickl M & Ries CH Comparison of 3D and 2D tumor models reveals enhanced HER2 activation in 3D associated with an increased response to trastuzumab. Oncogene 28:461-8 (2009). IP ; Human . Read more (PubMed: 18978815) »

See 1 Publication for this product

Product Wall

Thank you for your enquiry. Please see the information below from the originator of the Affibody. The application note we have published on Immunoprecipitation using the Anti-HER2 Affibody(r) molecule is recommending to boil the protein in sample b...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"