Lgt agarose type 7

Manufactured by Merck Group

Sourced in Germany

LGT agarose type VII is a laboratory equipment product manufactured by Merck Group. It is a type of agarose, a polysaccharide derived from seaweed, commonly used as a gel matrix in various laboratory techniques, such as electrophoresis and immunoassays. The core function of LGT agarose type VII is to provide a stable and consistent gel matrix for the separation and analysis of macromolecules, such as proteins and nucleic acids.

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Lab products found in correlation

Polyclonal rabbit anti human vwf antibody, by Agilent Technologies (1 mentions)

2 protocols using lgt agarose type 7

SDS-agarose Electrophoresis for VWF Multimers

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SDS-agarose discontinuous gel electrophoresis was performed as previously described [12] (link). Briefly, medium resolution (1.6%) gel (LGT agarose type VII, Sigma, Munich, Germany) was prepared and the samples were diluted according to their VWF content in sample buffer. Electrophoresis was performed for ∼18 h at 55 V. After electrophoresis, the multimers were transferred onto nitrocellulose membrane by electroblotting using transfer buffer (0.05 M phosphate, pH 7.4 with 0.04 M SDS, without methanol). After transfer, nonspecific binding sites were blocked with low-fat milk. All of the following incubation and washing steps were performed in low-fat milk. The membrane was incubated overnight at a 1∶2000 dilution of polyclonal rabbit anti-human VWF antibody (Dako, Glostrup, Denmark) or 1∶250 dilution of monoclonal mouse anti-human AAT type PiZ antibody (Clone ATZ11). Detection was performed using chemiluminescence (Fluorchem™, Alpha Innotech Corp., San Leandro, California).

Goltz D., Hittetiya K., Yadegari H., Driesen J., Kirfel J., Neuhaus T., Steiner S., Esch C., Bedorf J., Hertfelder H.J, & Fischer H.P. (2014). ATZ11 Recognizes Not Only Z-α1-Antitrypsin-Polymers and Complexed Forms of Non-Z-α1-Antitrypsin but Also the von Willebrand Factor. PLoS ONE, 9(3), e91538.

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VWF-Multimer Analysis in von Willebrand Disease

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VWF-multimers were analyzed in normal human pooled plasma (NHPP; CRYOcheck, Canada) and compared to Type 3 VWD (absent VWF) and Type 2A VWD (reduced VWF) patients as described previously.^{45 (link)} Briefly, VWF-multimers were separated by sodium dodecylsulfate–agarose electrophoresis, transferred onto a nitrocellulose membrane, detected with anti-human VWF antibody–horseradish peroxidase (DAKO, Glostrup, Denmark), and visualized by luminescence.
In addition, cell lysates of BOECs from Type 3 VWD were examined for VWF-multimers (Figure 2b). Multimer analysis was performed as described before^{46 (link)} in gels of medium resolution (1.6% LGT agarose type VII, Sigma-Aldrich, Munich, Germany). Gel analysis was performed using software provided with the video detection system (AlphaEaseFC Stand Alone software, Alpha Innotech Corp., San Leandro, CA). Samples with the same quantity of VWF:Ag were applied to the gels. The sensitivity of the assay is high compared to that of the VWF:Ag enzyme-linked immunosorbent assay (detection limit 0.038 U/ml = ~2 pg compared to 0.08 U/ml = ~4 pg).

Noone D.G., Riedl M., Pluthero F.G., Bowman M.L., Liszewski M.K., Lu L., Quan Y., Balgobin S., Schneppenheim R., Schneppenheim S., Budde U., James P., Atkinson J.P., Palaniyar N., Kahr W.H, & Licht C. (2016). Von Willebrand factor regulates complement on endothelial cells. Kidney international, 90(1), 123-134.

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