By GE HEalthcare

Show description

Read or Download affinity chromatography principles and methods PDF

Best chemistry books

Alkaloids: Chemical and Biological Perspectives, Vol. 15

Acronycine, a powerful antitumor agent, was once came across within the bark of the small Australian Rutaceous tree, Acronychia baueri Schott. This new paintings provides a finished survey of the isolation, constitution selection, equipment of synthesis, and the organic homes of acronycine, in addition to an account of usual and artificial analogues of acronycine, and their organic homes.

Chemistry of Heterocyclic Compounds: Imidazole and Its Derivatives, Part I, Volume 6

Content material: bankruptcy I normal homes and constitution of the Imidazoles (pages 3–31): bankruptcy II The Alkyl? and Arylimidazoles (pages 33–54): bankruptcy III The Oxo? and Hydroxyimidazoles and their Sulfur Analogues (pages 55–110): bankruptcy IV The Halogenoimidazoles (pages 111–125): bankruptcy V The Nitro? , Arylazo?

Additional resources for affinity chromatography principles and methods

Example text

Strong affinity for monoclonal mouse IgG1 and rat IgG. Complete kit contains HiTrap Protein G HP (1 x 1 ml), accessories, pre-made buffers for 10 purifications and detailed experimental protocols. Protein G Sepharose 4 Fast Flow Supplied as a suspension ready for column packing. Human IgG, > 20 mg/ml medium 400 cm/h* Cow IgG, 23 mg/ml medium Goat IgG, 19 mg/ml medium Guinea pig IgG, 17 mg/ml medium Mouse IgG, 10 mg/ml medium Rat IgG, 7 mg/ml medium * See Appendix 4 to convert linear flow (cm/h) to volumetric flow rate.

4. Wash with at least 10 column volumes of binding buffer or until no material appears in the eluent, as monitored at A280. 5. Elute the IgY with 10 column volumes of elution buffer. 6. Wash the column with 8 column volumes of wash buffer. 7. Immediately re-equilibrate the column with 5 column volumes of binding buffer. 8 M Na2SO4. The sample should have the same concentration of Na2SO4 as the binding buffer. An increase in salt concentration will reduce the purity of the eluted IgY. The purity of the eluted IgY may be improved by using gradient elution with, for example, a linear gradient 0–100% elution buffer over 10 column volumes, followed by elution with 100% elution buffer for a few column volumes.

4. Equilibrate the column with 10 column volumes of binding buffer. 5. Apply sample at a flow rate 1–4 ml/min (1 ml column) or 5 ml/min (5 ml column). Collect the flow-through fraction. A pump is more suitable for application of sample volumes greater than 15 ml. 6. Wash with 10 column volumes of binding buffer. Collect wash fraction. 7. Elute with 5 column volumes of elution buffer. Collect eluted fractions in small fractions such as 1 ml to avoid dilution of the eluate. 8. Wash with 10 column volumes of binding buffer.

Download PDF sample

Rated 4.16 of 5 – based on 8 votes