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Hitrap igm purification hp

Manufactured by GE Healthcare
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The HiTrap IgM Purification HP is a lab equipment product developed by GE Healthcare. It is designed for the purification of immunoglobulin M (IgM) from a variety of biological samples. The product utilizes a prepacked column format to facilitate the purification process.

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Lab products found in correlation

Alexa fluor 594 nhs ester, by Thermo Fisher Scientific (1 mentions) Pristane, by Merck Group (1 mentions) Mabtrap kit, by GE Healthcare (1 mentions) Histrap excel 5 ml column, by GE Healthcare (1 mentions) Hitrap protein a 5 ml column, by GE Healthcare (1 mentions) Superose 6 increase 10 300 gl, by GE Healthcare (1 mentions) Superdex 200 column, by GE Healthcare (1 mentions)

Close spelling variants of this product

HiTrap™ IgM Purification HP column

3 protocols using hitrap igm purification hp

1

Immunohistochemical Profiling of Muscle Fiber Types

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Gastrocnemius muscles were freshly frozen in isopentane cooled in liquid nitrogen, after which transverse 20-μm cryostat sections were used for muscle immunohistochemistry and morphological analysis. After mounting on MAS-coated slides (Matsunami glass), the sections were fixed in cold acetone and processed for immunohistochemical staining as described above using mouse monoclonal antibodies to the type I, IIA and IIB myosin heavy chain (MyHC) isoforms (HB287, HB277 and HB283, respectively; ATCC). Hybridoma clones HB287 and HB277 were intraperitoneally injected into nude mice, and the ascites fluid was harvested and labeled with Alexa Fluor594 NHS Ester (Molecular Probes). For HB283, the IgM fraction was purified from culture supernatant using HiTrap IgM Purification HP (GE), and Alexa Fluor594-labeled donkey anti-mouse IgM, μ Chain Specific (Jackson ImmunoResearch Laboratories) was used as a secondary antibody for detection. For morphological analysis, cryosections were stained with hematoxylin and eosin.
Morisaki Y., Niikura M., Watanabe M., Onishi K., Tanabe S., Moriwaki Y., Okuda T., Ohara S., Murayama S., Takao M., Uchida S., Yamanaka K, & Misawa H. (2016). Selective Expression of Osteopontin in ALS-resistant Motor Neurons is a Critical Determinant of Late Phase Neurodegeneration Mediated by Matrix Metalloproteinase-9. Scientific Reports, 6, 27354.
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2

Monoclonal Antibody Purification Protocol

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In order to obtain large concentrations of mAbs, for further studies, ascitic fluids of monoclonal antibodies BURK24 and BURK37 were generated produced in 2,6,10,14-tetramethyl pentadecane i.e., Pristane (Sigma, India) primed retired breeder BALB/c mice, by injecting BURK24 and BURK37 hybridomas as described by R B Westerman et al.[33] (link) and purified using mAbTrap kit and HiTrap IgM Purification HP (GE Healthcare, UK) respectively as per the manufacturer's instructions. The purified mAbs were lyophilized, quantified and reconstituted in sterile 1× PBS according to the required concentrations of each study.
Peddayelachagiri B.V., Paul S., Makam S.S., Urs R.M., Kingston J.J., Tuteja U., Sripathy M.H, & Batra H.V. (2014). Functional Characterization and Evaluation of In Vitro Protective Efficacy of Murine Monoclonal Antibodies BURK24 and BURK37 against Burkholderia pseudomallei. PLoS ONE, 9(3), e90930.
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3

Engineered Nanobody-Fc Antibody Constructs

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The dimer constructs (DR14 and DS43) were engineered via fusing nanobody to hFc. The homo-trimer versions of R14 and S43 (TR14 and TS43) were constructed via head-to-tail with (GGGGS)3 linkers and a C-terminal His-tag. The coding sequence of R14 or S43 fused to the Fc of human IgM was cloned into the pCAGGS vector to generate IgM versions of the antibodies (MR14 and MS43). The coding sequence of TR14 was synthesized by Nanjing GenScript Biotech Co., Ltd., and TS43 was synthesized by Tsingke Biotechnology Co., Ltd. DR14 and DS43 were expressed in Freestyle 293F cells and were purified using a HiTrap Protein A 5-mL column (GE Healthcare) and Superdex 200 column (GE Healthcare). TR14 and TS43 were expressed in Freestyle 293F cells and were purified using a HisTrap EXCEL 5-mL column (GE Healthcare) and Superdex 200 column (GE Healthcare). The recombinant plasmid for MR14 or MS43 and a human J-chain expressing vector were co-transfected into Freestyle 293F cells to express the MR14 or MS43 proteins, which were further purified by HiTrap™ IgM Purification HP (GE Healthcare) and Superose 6 Increase 10/300 GL (GE Healthcare) chromatography.
Liu H., Wu L., Liu B., Xu K., Lei W., Deng J., Rong X., Du P., Wang L., Wang D., Zhang X., Su C., Bi Y., Chen H., Liu W.J., Qi J., Cui Q., Qi S., Fan R., Jiang J., Wu G., Gao G.F, & Wang Q. (2023). Two pan-SARS-CoV-2 nanobodies and their multivalent derivatives effectively prevent Omicron infections in mice. Cell Reports Medicine, 4(2), 100918.
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