Overview

  • Product name
    10kD Spin Column
  • Product overview

    The 10kD Spin Columns (ab93349) are disposable ultrafiltration devices for the concentration of biological samples. They can be used in either a swing bucket or fixed angle rotors which accept 2.0 mL centrifuge tube at maximum speed 10000 x g (5-10 mins). The vertical membrane design and thin channel filtration chamber minimizes membrane fouling and provides high speed concentration, even laden solutions. It is specifically designed with low internal surface and membrane area in order to achieve superior recovery from dilute solution.


    Maximum initial sample volume is 500µL. The dead stop volume is approximately 20-25µL


    Visit our FAQs page for tips and troubleshooting.

  • Tested applications
    Suitable for: Purificationmore details

Properties

Applications

Our Abpromise guarantee covers the use of ab93349 in the following tested applications.

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

Application Abreviews Notes
Purification Use at an assay dependent concentration.

Images

References

This product has been referenced in:
  • Li FL  et al. Acetylation accumulates PFKFB3 in cytoplasm to promote glycolysis and protects cells from cisplatin-induced apoptosis. Nat Commun 9:508 (2018). Read more (PubMed: 29410405) »
  • Riesberg LA  et al. Creatinine downregulates TNF-a in macrophage and T cell lines. Cytokine 110:29-38 (2018). Read more (PubMed: 29698843) »
See all 12 Publications for this product

Customer reviews and Q&As

1-10 of 30 Abreviews or Q&A

Answer



It is hard to predict how long will be required to spin the volume through the spin filter. It depends on how much protein is in the solution to be filtered as higher protein concentrations require longer spin times. I recommend spinning for 5 min at 4 deg C at first and then check to see how much volume has gone through. Then you will have a good idea as to how much longer will be required to push the remainder through.

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Answer

Thank you for contacting us.

Always ensure that you do not load more than 450 µl of diluted, non-viscous samples. I understand in the case of plasma you may not wish to dilute further. If you still have a difficulty in recovering a reasonable volume back, please centrifuge the samples for ˜30-45 min at 12000 rpm at 4°C.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer



Ich kann bestätigen, dass die "spin cloumns" verwendet werden um alles, das grösser ist als 10kDa loszuwerden.

Bitte verwenden Sie den deshalb Durchfluss im ATP assay kit.

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Answer



The columns have been tested to ensure that the samples are separated correctly. The fragments have been tested using several of our assay kits, verifying that analytes above/below 10 kDa are present in the correct fragment.

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Answer

You are exactly correct, insoluble material, whether from cells or tissue, needs to be removed prior to loading into the 10kDa spin column. Thus, after lysis/homogenization, centrifuge your sample to pellet the insoluble material. Then take the supernatant as your sample and load into the 10kDa spin column.

Typically, an equivalent volume of sample will be loaded per well as the assay measures the amount of lactate present, but please refer to your lactate assay kit protocol. One of our lactate assay protocols is provided through the link below and uses an equivalent volume of sample per well:
https://www.abcam.com/ps/products/65/ab65331/documents/ab65331%20L%20Lactate%20Assay%20Kit%20Colorimetric%20(Website).pdf

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Answer

We recommend the use of the 10 kDa filters irrespective of your immediate or later usage of the lysed samples. The filters help in separating out proteins which can chew up the analyte. You most likely will not get efficient results without the fliters.

We carry 10 kD spin columns to assist in using this kit. The link for these columns is given below:
https://www.abcam.com/10kd-spin-column-ab93349.html

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Answer

Thanks for your response.
I strongly recommend sticking to the kit protocol. Adding solutions (such as ethanol and perchloric acid) other than those provided in the kit will compromise the efficiency of lactate detection. The centrifugation step can be used to remove bubbles after homogenization in the assay buffer.
I hope this response proves helpful. Please do not hesitate to reply or contact us with any additional questions.

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Answer

Thank you for contacting Abcam.
Since the lactate dehydrogenase (LDH) in tissue lysates degrades lactate, be sure to keep your samples at -80ºC. Below is our recommended homogenization protocol:
1. Homogenize the tissue samples (should contain at least 2 million cells) in the assay buffer that is provided for 10-50 passes on ice.
2. Centrifuge the sample and collect the supernatant.
3. Load the supernatant onto a 10kda spin column (ab93349 https://www.abcam.com/10kd-spin-column-ab93349.html) for deproteinization to remove LDH. Use the eluate for your subsequent assays.
To make sure that your readings from these samples are within the standard curve, we suggest using several doses of your sample when running your assay.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
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Answer

Thank you for contacting us.

Yes, you can definitely use 10 kD spin filter to filter out all the proteins from samples. We routinely use and recommend these filters in conjugation with many of our assays kits.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Question
Answer

Merci de nous avoir contactés.

Nous suggérons de mettre votre surnageant (maximum 500 ul) dans la colonne à une vitesse de 10000 x g pour 5 minutes. Par contre les temps varient donc il se peut que 5 minutes ne suffisent pas et qu’une centrifugation plus longue soit nécessaire.

J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

Vous utilisez nos produits? Partagez vos résultats sous forme d'Abreview et recevez en échange des CADEAUX!
https://www.abcam.com/abreviews

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1-10 of 30 Abreviews or Q&A

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

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