Preparing a SDS-PAGE Gel - Revision history

Reddj: added SOP requirement-JR

2020-06-04T16:38:03Z

added SOP requirement-JR

← Older revision		Revision as of 16:38, 4 June 2020
Line 1:		Line 1:
			==SOP==
			*[[SOP- Electrophoresis]]

	==Separating Gel==		==Separating Gel==
	*For 12 mL final volume (2 gels)		*For 12 mL final volume (2 gels)

Davebrid at 12:08, 5 August 2009

2009-08-05T12:08:30Z

← Older revision		Revision as of 12:08, 5 August 2009
Line 16:		Line 16:
	\| TEMED \|\| 30 uL \|\| 30 uL \|\| 30 uL \|\| 30uL		\| TEMED \|\| 30 uL \|\| 30 uL \|\| 30 uL \|\| 30uL
	\|}		\|}
	*For a Phos-Tag Gel, add 24 uL 5 mM Phos-Tag and 24 uL 10 mM MnCl in the separating gel only		*For a [[Phos-Tag Electrophoresis and Western Blotting\|Phos-Tag Gel]], add 24 uL 5 mM Phos-Tag and 24 uL 10 mM MnCl in the separating gel only

	==Stacking Gel==		==Stacking Gel==
Line 39:		Line 39:
	[[Category: SDS-PAGE]]		[[Category: SDS-PAGE]]
	[[Category: Electrophoresis]]		[[Category: Electrophoresis]]
			[[Category: PhosTag]]
			[[Category: Phosphorylation]]

Davebrid at 12:07, 5 August 2009

2009-08-05T12:07:01Z

← Older revision		Revision as of 12:07, 5 August 2009
Line 16:		Line 16:
	\| TEMED \|\| 30 uL \|\| 30 uL \|\| 30 uL \|\| 30uL		\| TEMED \|\| 30 uL \|\| 30 uL \|\| 30 uL \|\| 30uL
	\|}		\|}
	*For a Phos-Tag Gel, add 24 uL 5 mM Phos-Tag and 24 uL 10 mM MnCl		*For a Phos-Tag Gel, add 24 uL 5 mM Phos-Tag and 24 uL 10 mM MnCl in the separating gel only

	==Stacking Gel==		==Stacking Gel==
Line 32:		Line 32:
	\| Water \|\| 4 mL		\| Water \|\| 4 mL
	\|-		\|-
	\| APS (make fresh, 0.1g/mL in water) ~~\|\| 200 uL \|\| 200 uL~~ \|\| 200 uL		\| APS (make fresh, 0.1g/mL in water) \|\| 200 uL
	\|-		\|-
	\| TEMED \|\| 30 uL ~~\|\| 30 uL \|\| 30uL~~		\| TEMED \|\| 30 uL
	\|}		\|}

	[[Category: SDS-PAGE]]		[[Category: SDS-PAGE]]
	[[Category: Electrophoresis]]		[[Category: Electrophoresis]]

Davebrid: added 6% gel

2009-08-05T12:06:31Z

added 6% gel

← Older revision		Revision as of 12:06, 5 August 2009
Line 4:		Line 4:
	*Pour into cassette, overlay with the top layer of water saturated butanol		*Pour into cassette, overlay with the top layer of water saturated butanol
	{\| border="1"		{\| border="1"
	! Component \|\| 7.5% \|\| 10% \|\| 12.5%		! Component \|\| 6% \|\|7.5% \|\| 10% \|\| 12.5%
	\|-		\|-
	\| [[SDS-PAGE Separating Gel Buffer]] \|\| 3 mL \|\| 4 mL \|\| 5 mL		\| [[SDS-PAGE Separating Gel Buffer]]\|\| 3 mL \|\| 3 mL \|\| 3 mL \|\| 3 mL
	\|-		\|-
	\| [[Acrylamide Solution]] \|\| 3 mL \|\| 3 mL \|\| 3 mL		\| [[Acrylamide Solution]]\|\| 2.4 mL \|\| 3 mL \|\| 4 mL \|\| 5 mL
	\|-		\|-
	\| Water \|\| 6 mL \|\| 5 mL \|\| 4mL		\| Water \|\| 6.6 mL\|\| 6 mL \|\| 5 mL \|\| 4mL
	\|-		\|-
	\| APS (make fresh, 0.1g/mL in water) \|\| 200 uL \|\| 200 uL \|\| 200 uL		\| APS (make fresh, 0.1g/mL in water) \|\| 200uL \|\| 200 uL \|\| 200 uL \|\| 200 uL
	\|-		\|-
	\| TEMED \|\| 30 uL \|\| 30 uL \|\| 30uL		\| TEMED \|\| 30 uL \|\| 30 uL \|\| 30 uL \|\| 30uL
	\|}		\|}
			*For a Phos-Tag Gel, add 24 uL 5 mM Phos-Tag and 24 uL 10 mM MnCl

	==Stacking Gel==		==Stacking Gel==

Davebrid: Created page with '==Separating Gel== For 12 mL final volume (2 gels) Combine, adding APS and TEMED last *Pour into cassette, overlay with the top layer of water saturated butanol {| border="1" !...'

2009-08-03T18:52:50Z

Created page with '==Separating Gel== *For 12 mL final volume (2 gels) *Combine, adding APS and TEMED last *Pour into cassette, overlay with the top layer of water saturated butanol {| border="1" !...'

New page

==Separating Gel==
*For 12 mL final volume (2 gels)
*Combine, adding APS and TEMED last
*Pour into cassette, overlay with the top layer of water saturated butanol
{| border="1"
! Component || 7.5% || 10% || 12.5%
|-
| [[SDS-PAGE Separating Gel Buffer]] || 3 mL || 4 mL || 5 mL
|-
| [[Acrylamide Solution]] || 3 mL || 3 mL || 3 mL
|-
| Water || 6 mL || 5 mL || 4mL
|-
| APS (make fresh, 0.1g/mL in water) || 200 uL || 200 uL || 200 uL
|-
| TEMED || 30 uL || 30 uL || 30uL
|}

==Stacking Gel==
*Rinse polymerized separating gel with water
*For 10 mL (2 gels)
*Combine, adding APS and TEMED last
*Pour over separating gel and insert comb
{| border="1"
! Component ||
|-
| [[SDS-PAGE Stacking Gel Buffer]]|| 2.5 mL
|-
| [[Acrylamide Solution]] || 1.7 mL
|-
| Water || 4 mL
|-
| APS (make fresh, 0.1g/mL in water) || 200 uL || 200 uL || 200 uL
|-
| TEMED || 30 uL || 30 uL || 30uL
|}

[[Category: SDS-PAGE]]
[[Category: Electrophoresis]]

Preparing a SDS-PAGE Gel - Revision history

Reddj: added SOP requirement-JR

Davebrid at 12:08, 5 August 2009

Davebrid at 12:07, 5 August 2009

Davebrid: added 6% gel

Davebrid: Created page with '==Separating Gel== *For 12 mL final volume (2 gels) *Combine, adding APS and TEMED last *Pour into cassette, overlay with the top layer of water saturated butanol {| border="1" !...'

Davebrid: Created page with '==Separating Gel== For 12 mL final volume (2 gels) Combine, adding APS and TEMED last *Pour into cassette, overlay with the top layer of water saturated butanol {| border="1" !...'