# Protein Isolation for Western Blots (protocols)?

**URL:** <https://forum.painresearcher.net/t/protein-isolation-for-western-blots-protocols/785>\
**Category:** Experimental Techniques\
**Created:** [January 10, 2021, 10:50pm UTC](https://forum.painresearcher.net/t/protein-isolation-for-western-blots-protocols/785 "2021-01-10T22:50:44Z")\
**Posts on this page:** 4\
**Page:** 1

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**Author:** ![achamess](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/achamess/32/534_2.png) [@achamess](https://forum.painresearcher.net/u/achamess)\
**Post date:** [January 10, 2021, 10:50pm UTC](https://forum.painresearcher.net/t/protein-isolation-for-western-blots-protocols/785/1 "2021-01-10T22:50:44Z")

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Hey all,  
Looking to do some protein work soon. I have some personal protocols from grad school but it’d be nice to see what people are doing.

If you’re willing, share your protocol for:

**Mouse**

- Spinal cord
- DRG
- Nerve

**Human**

- DRG
- Nerve

I made video a few years back on Mouse DRG lysate prep based on the protocol @tberta taught me

[![](https://canada1.discourse-cdn.com/flex030/uploads/painresearcher/original/1X/aa8c37d6adad662b61c684993051c48822b6bb67.jpeg "Dorsal Root Ganglia (Mouse) Protein Lysate Preparation - Translational Pain Research Lab Protocols") ](https://www.youtube.com/watch?v=Su3qVht1lBQ)

I used to use a BeadBeater as well for many things. I like to avoid the sonicator whenever possible, but I don’t have the bead beater anymore.

@tberta @sshiers @CandlerPaige @thicunha @ShanTan

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**Author:** ![achamess](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/achamess/32/534_2.png) [@achamess](https://forum.painresearcher.net/u/achamess)\
**Post date:** [January 18, 2021, 4:24pm UTC](https://forum.painresearcher.net/t/protein-isolation-for-western-blots-protocols/785/2 "2021-01-18T16:24:03Z")

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Hey all. I’ll answer my own question. I used a Dounce homogenizer for hDRG and hSC, and  
[https://www.dwk.com/na/kimble-kontes-duall-micro-tissue-grinder-02-ml-885470-0000](https://www.dwk.com/na/kimble-kontes-duall-micro-tissue-grinder-02-ml-885470-0000) for mouse DRG and nerve.

RIPA + complete mini + halt protease inhibitor

Looked at TUBB3 by capillary electrophoresis immunoassay (like a Western).

Looks good! Can’t say that all proteins were extracted optimally but it’s good enough to get some protein.

 ![image](https://canada1.discourse-cdn.com/flex030/uploads/painresearcher/original/1X/7376f43a9c8d0809186123b7c05889dddb9e363a.png)

I’ll post protocols soon.

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**Author:** ![bdagostin](https://avatars.discourse-cdn.com/v4/letter/b/f1d935/32.png) [@bdagostin](https://forum.painresearcher.net/u/bdagostin)\
**Post date:** [July 19, 2024, 5:03pm UTC](https://forum.painresearcher.net/t/protein-isolation-for-western-blots-protocols/785/3 "2024-07-19T17:03:33Z")

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Greetings,  
For mouse DRG - would you pool multiple DRG together to have enough protein to extract? If so, how many? I was hoping that I would be able to extract enough protein from perhaps L4 and L5 DRG from a few animals, but worry about low protein concentration. Also would you mind sharing your protocol?

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**Author:** ![aloegum](https://avatars.discourse-cdn.com/v4/letter/a/a6a055/32.png) [@aloegum](https://forum.painresearcher.net/u/aloegum)\
**Post date:** [July 30, 2024, 6:12pm UTC](https://forum.painresearcher.net/t/protein-isolation-for-western-blots-protocols/785/4 "2024-07-30T18:12:41Z")

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We pooled multiple mouse Lumbar DRGs (6 pairs ) and lysed in 200 uL RIPA+0.5% SDS buffer. We usually obtain 300 ug total protein.