ACSF Media - Revision history

Davebrid: changed category tag

2017-08-02T14:28:05Z

changed category tag

← Older revision		Revision as of 14:28, 2 August 2017
Line 48:		Line 48:
	Add ~450 mL water, bring pH to 7.4. Bring final volume up to 500 mL and sterile filter into an autoclaved bottle. Store at 4C.		Add ~450 mL water, bring pH to 7.4. Bring final volume up to 500 mL and sterile filter into an autoclaved bottle. Store at 4C.

	[[ Category: ~~Buffers~~ ]]		[[ Category: Buffer ]]
	[[ Category: Seahorse ]]		[[ Category: Seahorse ]]
	[[ Category: Cell Culture ]]		[[ Category: Cell Culture ]]

Davebrid: Updated KHPO4 concentration and fixed a typo

2017-04-27T13:21:28Z

Updated KHPO4 concentration and fixed a typo

← Older revision		Revision as of 13:21, 27 April 2017
Line 1:		Line 1:
	This media stock is the media for both the mitochondrial stress assays for skeletal muscle fibers. Because it is fairly well buffered, ECAR results are not reliable.		This media stock is the media for both the mitochondrial stress assays for skeletal muscle fibers. Because it is fairly well buffered, ECAR results are not reliable.
	Recheck the pH after warming and prior to each use. These volumes are to make ''500 mL'' of media:
			This recipe and protocol for isolating/measuring muscle fibers is found [http://www.agilent.com/cs/pubimages/misc/TB_Intact_Muscle_Fibers.pdf here]
			Recheck the pH after warming and prior to each use. These volumes are to make '''500 mL''' of media:

	{\| class="wikitable"		{\| class="wikitable"
Line 18:		Line 20:
	\| 3.5 mM		\| 3.5 mM
	\|-		\|-
	\| Calcium ~~Chloride~~		\| Calcium chloride
	\| 1 M		\| 1 M
	\| 650 uL		\| 650 uL
	\| 1.3 mM		\| 1.3 mM
	\|-
	~~\| Potassium chloride~~
	~~\| 1 M~~
	~~\| 2.7 mL~~
	~~\| 5.4 mM~~
	\|-		\|-
	\| Potassium phosphate		\| Potassium phosphate
	\| 1M		\| 0.2 M
	\| ~~200 uL~~		\| 1 mL
	\| 0.4 mM		\| 0.4 mM
	\|-		\|-

Davebrid: Added categories

2017-04-27T12:41:59Z

Added categories

← Older revision		Revision as of 12:41, 27 April 2017
Line 50:		Line 50:

	Add ~450 mL water, bring pH to 7.4. Bring final volume up to 500 mL and sterile filter into an autoclaved bottle. Store at 4C.		Add ~450 mL water, bring pH to 7.4. Bring final volume up to 500 mL and sterile filter into an autoclaved bottle. Store at 4C.

			[[ Category: Buffers ]]
			[[ Category: Seahorse ]]
			[[ Category: Cell Culture ]]
			[[ Category: Metabolism ]]
			[[ Category: Respiration ]]
			[[ Category: Tissue Culture ]]
			[[ Category: Mouse Tissues ]]
			[[ Category: Muscle Cells ]]

Davebrid: Wrote initial table for buffers

2017-04-27T12:39:29Z

Wrote initial table for buffers

New page

This media stock is the media for both the mitochondrial stress assays for skeletal muscle fibers. Because it is fairly well buffered, ECAR results are not reliable.
Recheck the pH after warming and prior to each use. These volumes are to make ''500 mL'' of media:

{| class="wikitable"
! style="font-weight: bold;" | Reagent
! style="font-weight: bold;" | Stock Concentration
! style="font-weight: bold;" | Volume (500 mL)
! style="font-weight: bold;" | Final Concentration
|-
| Sodium chloride
| 4 M
| 15 mL
| 120 mM
|-
| Potassium chloride
| 1 M
| 1.75 mL
| 3.5 mM
|-
| Calcium Chloride
| 1 M
| 650 uL
| 1.3 mM
|-
| Potassium chloride
| 1 M
| 2.7 mL
| 5.4 mM
|-
| Potassium phosphate
| 1M
| 200 uL
| 0.4 mM
|-
| Magnesium chloride
| 1 M
| 0.5 uL
| 1 mM
|-
| HEPES
| 1 M
| 2.5 mL
| 5 mM
|-
| D-Glucose
| Powder
| 1.35 g
| 15 mM
|}

Add ~450 mL water, bring pH to 7.4. Bring final volume up to 500 mL and sterile filter into an autoclaved bottle. Store at 4C.