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What is the size of the agarose beads?
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ANSWER: |
Thank you for your enquiry. The bead size is 60-160um. If you have any further questions then please get back in touch with me.
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We purchased your Anti-IgG ab 7018. 1) I would like to know what does mean: Sufficient antibody capacity is provided to bind a minimum of 5mg of pure mouse IgG (Abcam instruction for Anty-IgG ab 7018, Application notes point) 2) I would like to use your product in cartridge (column) for purification of mouse antibodies. About 1000 ng of the first antibody will be present in my sample. What amount of your Anti-IgG matrice I should use? 3) What is the bounding density of Anti-IgG ab 7018? |
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ANSWER: |
Thank you for your enquiry. We can confirm that 20mg of antibody can bind at least 5mg mouse IgG. Theoretically, it requires 0.04uL of agarose bead to purify 1000ng of antibody. We are very sorry but bounding density is not available, our apologies for the inconvenience. |
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i'm interested in your anti mouse agarose conjugated ab 7018 but i need to know the exact binding capacity
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ANSWER: |
It really depends on how to use it and what to use it for. For an optimized purification system, to purify IgG, the binding capacity is about 1-2 mg of IgG per mL of agarose resin. |
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I reviewed your posted Q&A regarding the procedure for purifying murine monoclonal antibodies. I have been washing with Tris-Saline Buffer pH 8.0 followed by elution with 3M KSCN. This is the same protocol I have been using for Protein A and G, but my yield when using your column has been very small. (I even cycle and incubate my sample in the column before I wash) Is there a reason for this, and how can I increase my yield? Thank you for your time. |
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ANSWER: |
We suggest that you try PBS pH 7.2 as washing buffer and 0.1M Glycine pH 2.5 as elution buffer. |
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Dear Sir or Madam, Our research is related to purification of murine monoclonal antibody from the hybridoma cell culture medium. We tried several methods, such as Protein A and Protein G. The results showed that the IgG was contaminated with bovine IgG from serum in the medium. We talked with other experienced person around. They recommended the anti-mouse IgG agarose. Finally, we found this product, ab7018 from your company. But we are not pretty sure if it will work or not for us. Could you please specify the protocol which can just purify murine monoclonal antibody from the cell culture with this product? Is there any special procedures we need to apply? Thank you in advance. Looking forward to your response. |
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ANSWER: |
Yes, this agarose bead conjugated antibody can be used for the affinity purification as stated. The protocol would be a general protocol of affinity purification. Simply load the column with the Mouse monoclonal antibody under the following conditions: basic to neutral pH and a high salt concentration, then elute it with a low pH and low salt concentration. |
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