Dear friend Sarah L Fox

Hey there Sarah L Fox! I am here, ready to tackle your Western blot challenges. Now, histone modifications on H3 can indeed make it a tricky loading control. If H3 and H4 are not cutting it and Ponceau isn't playing nice, let's think outside the PVDF membrane!

Considering the inconsistencies with H4, lamin B might be worth a shot. Lamin B is a nuclear envelope protein and could serve as an alternative loading control, especially if you're working with nuclear extracts.

Here's a quick guide:

1. **Optimize Extraction:**

- Ensure your histone extraction is top-notch using the abcam kit. Proper extraction can save you Sarah L Fox from loading control woes.

2. **Membrane Type:**

- Since Ponceau isn't doing the trick, consider trying a nitrocellulose membrane. Sometimes, different membranes have different affinities.

3. **Lamin B as a Control:**

- Lamin B has been used as a loading control in some cases, especially in studies involving nuclear proteins. It's worth trying, but always validate with literature or pilot experiments.

4. **PVDF Stripping Woes:**

- If you're having trouble stripping the PVDF membrane, consider trying a fresh one for a cleaner start.

5. **Alternate Controls:**

- If lamin B doesn’t pan out, consider beta-actin or GAPDH. While they're not without their own challenges, they are commonly used and might suit your needs.

6. **Trial and Error:**

- Sometimes, it's all about experimenting. Tweak concentrations, try different gels, and play with transfer conditions. The perfect loading control might just be waiting for you Sarah L Fox to discover it.

7. **Consult the Literature:**

- Before you Sarah L Fox dive too deep, check recent literature. Others in your field might have encountered similar challenges and found solutions.

Remember, science is an adventure, and the best protocols often come from a bit of exploration. Let me know how it goes, and we'll navigate this Western blot wilderness together Sarah L Fox!

Kaushik Shandilya thank you! Read my mind.. the blots are currently in lamin B but im worried it wont work as we used the histone extraction kit from abcam and unsure if anyything but histones will be present!

can we use GAPDH if we use histone extraction kit?! I detect a similar issue.

I think the next step is to use nitrocellulose membranes. thank you! see how lamin B goes tomorrow :D