Author Topic: White line in the acidic end of the strips  (Read 2837 times)

Violeta G

  • Newbie
  • *
  • Posts: 9
White line in the acidic end of the strips
« on: May 28, 2013, 05:35:36 AM »
Hello!

I've been getting an extrange white line near the acidic end of all my strips (Immobiline DryStrips, GE Healthcare, pH 4-7). This produces a turbid, poorly focused region in the acidic side of the 2D gels.

My IEF conditions are: active rehydration (30V-6h; 60V-6h); 500V-1h; 1000V-1h; gradient to 8000V-30min; 8000V-8h. I've also tried a final step of 8000V-6.30h and 8000V-7.30h.

I work with pancreatic samples, specifically with a subcelular soluble fraction. I've changed anything since my first gels (in which this white line wasn't shown).

Could anyone give me a piece of advice? I'm running out of ideas.

Thanks. V.

Violeta G

  • Newbie
  • *
  • Posts: 9
Re: White line in the acidic end of the strips
« Reply #1 on: June 12, 2013, 10:42:19 AM »
I'd appreciate any idea you can give me! Thank you.

watdo

  • Newbie
  • *
  • Posts: 6
Re: White line in the acidic end of the strips
« Reply #2 on: September 24, 2013, 07:21:59 PM »
Are you usign cuploading with the manifold because I see this white line on mine where the cup sits. The smears you see are where the electrodes are?

Violeta G

  • Newbie
  • *
  • Posts: 9
Re: White line in the acidic end of the strips
« Reply #3 on: September 25, 2013, 05:18:40 AM »
Hi watdo!

I'm not using cuploading but in-gel rehydration (with strip holders). That smearing appears near the (+) electrode, but it spreads further.

Can you tell me what the composition of your rehydration buffer is?

watdo

  • Newbie
  • *
  • Posts: 6
Re: White line in the acidic end of the strips
« Reply #4 on: September 25, 2013, 05:08:24 PM »
I use GE DeStreak for rehydration with 0.5% IPG buffer added into it. The DeStreak contains urea, thiourea, CHAPS, and DeStreak Reagent. When I see a white line on mine it is where my gel has dried out.