Low Fluorescence Western Membrane (PVDF) (ab133411)

Reviews (1)Q&A (3)References (1)

Overview

  • Product name
    Low Fluorescence Western Membrane (PVDF)
    See all Western blot membrane reagents
  • Tested applications
    Suitable for: WBmore details
  • General notes

    Pre-cut membranes (10 sheets, 7 x 9 cm), compatible with gels 10 x 10 cm and smaller. Note: PVDF membranes must be activated prior to use. Membrane activation protocol: 1. Pre-wet the membrane in methanol for a few moments until the membrane is completely wet. 2. Transfer the wet membrane from methanol to purified water and incubate on a rocker or shaker for at least 5 minutes. Make sure that the membrane is fully immersed in water and does not float on the water’s surface. 3. Transfer the membrane from water to transfer buffer, and incubate for 5 minutes.

Properties

Applications

Our Abpromise guarantee covers the use of ab133411 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
WB Use at an assay dependent concentration.

References

This product has been referenced in:
  • Jebali R  et al. Lactobacillus plantarum alleviate aflatoxins (B1 and M1) induced disturbances in the intestinal genes expression and DNA fragmentation in mice. Toxicon 146:13-23 (2018). WB . Read more (PubMed: 29574215) »
See 1 Publication for this product

Publishing research using ab133411? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

PVDF membrane with Optiblot reagents works like charm with excellent results when developed

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
We tried other popular PVDF membranes and just didn't get the sort of clarity we wanted until we started using the Optiblot reagent kits. The PVDF is recommended for use with the Optiblot kits and we have be able to obtain excellent transfer of protein from 17-well and 12-well Optiblot gels onto the membranes as well subsequent detection of a weakly expressed membrane protein.

The image is taken straight after blotting for 2 and a half hour in the Optiblot reagents, followed by Ponceau S staining for 1 minute, with ladder in blue and 17-lanes of protein (70ug per lane). The developed images is for detection of Beta-Actin.
The membrane itself is quite robust to stripping and not prone to tearing.
Overall, compared to other PVDF membranes, when combined with the Optblot reagents, we have seen a successful series of blots and detection of proteins.

Dr. Raj Badhan

Verified customer

Submitted Jan 23 2014

Question

  Thank you very much for your information.   If I am transferring a small protein molecule around 3 kDa MW, what transfer condition would you recommend me to use in terms of voltage and time of transfer using this membrane?  

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Answer

I just noticed that I had forgotten to add the Western Blot guide that I mentioned in the last email. I have attached the guide to this email.

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

Thank you very much for your information.   If I am transferring a small protein molecule around 3 kDa MW, what transfer condition would you recommend me to use in terms of voltage and time of transfer using this membrane?   Thank you very much for your time,

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Answer

Thank you for contacting us.

Generally when transferring a very small protein, it is best to use a semi-dry transfer procedure. These usually run at 100V for 40-60 minutes. I would recommend transfer times at the lower end to this range. It is also helpful to remove SDS from your transfer buffer when transferring small proteins. I have included with this email our Western blot guide which includes further transfer information. Below I have included a link to our WB reagents page which has a semi-dry transfer buffer recipe. Finally, I have a brief protocol for semi-dry transfer which I will place at the end of this email.

https://www.abcam.com/index.html?pageconfig=resource&rid=13045

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



Semi-Dry Transfer

1. Cut 6-10 sheets of filter paper same size as the gel.
2. Cut membrane to the size of the gel.
3. Membrane activation for PVDF: soak in methanol for 15 seconds and then transfer to a containerfilled with distilled water for 5 minutes.
4. Soak membrane in semi-dry transfer buffer for 10 minutes while preparing transfer sandwich
5. Soak filter paper in transfer buffer for several minutes to 10 minutes to avoid trapping of air bubble.
6. Assemble sandwich in the following order:

3-5 sheets filter paper -

Gel

Membrane

3-5 sheets filter paper +



7. Use a roller to roll over the sandwich gently to remove trapped air bubbles.
8. Apply few ml transfer buffer on top of the sandwich to avoid drying out during membrane transformation.
9. Transfer at 100 V for 40-60 minutes.
10. Block and Stain the membrane as normal

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Question

Customer kindly contacted us regarding the pore size of the PVDF membranes (ab133411) as well as for further information about other products.

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Answer

Thank you for contacting us.

I have been in contact with the lab regarding ab133411. The pore size of the PVDF membrane is 0.45um

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

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