Seahorse XF Media - Revision history

Davebrid: changed category tag

2017-08-02T14:31:49Z

changed category tag

← Older revision		Revision as of 14:31, 2 August 2017
Line 47:		Line 47:
	Add ~450 mL water, bring pH to 7.4 carefully (there is very little buffering capacity, so add NaOH slowly). 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 carefully (there is very little buffering capacity, so add NaOH slowly). 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: added categories

2017-04-27T12:41:20Z

added categories

← Older revision		Revision as of 12:41, 27 April 2017
Line 46:		Line 46:

	Add ~450 mL water, bring pH to 7.4 carefully (there is very little buffering capacity, so add NaOH slowly). 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 carefully (there is very little buffering capacity, so add NaOH slowly). 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 ]]

Davebrid: updated stock concentrations

2017-03-10T14:07:13Z

updated stock concentrations

← Older revision		Revision as of 14:07, 10 March 2017
Line 1:		Line 1:
	This minimal media stock is the base media for both the glycolysis and mitochondrial stress assays. It contains no buffering and no carbon sources. It is a non-buffered media but still needs to be brought to pH 7.4 and checked before each assay.		This minimal media stock is the base media for both the glycolysis and mitochondrial stress assays. It contains no buffering and no carbon sources. It is a non-buffered media but still needs to be brought to pH 7.4 and checked before each assay.

	For glycolysis assays add:		For glycolysis assays add to warmed XF base media just before the assay and recheck pH:

	* 1-4 mM L-Glutamine		* 1-4 mM L-Glutamine (Generally 2 mM is 1mL/100mL of 200 mM stock)

	For mitochondrial assays add:		For mitochondrial assays and cellular phenotype assays add to warmed XF base media just before the assay and recheck pH:

	* 0.5-10 mM Sodium pyruvate		* 0.5-10 mM Sodium pyruvate (Generally 1 mM is 100uL/100mL of 1M stock)
	* 1-4 mM L-Glutamine		* 1-4 mM L-Glutamine (Generally 2 mM is 1mL/100mL of 200 mM stock)
	* 2-5 mM Glucose		* 2-25 mM Glucose (Generally 10 mM is 1mL/100mL of 1M stock)

	To make base media add the following:		To make base media add the following:

Davebrid: Wrote initial page

2017-02-22T21:07:47Z

Wrote initial page

New page

This minimal media stock is the base media for both the glycolysis and mitochondrial stress assays. It contains no buffering and no carbon sources. It is a non-buffered media but still needs to be brought to pH 7.4 and checked before each assay.

For glycolysis assays add:

* 1-4 mM L-Glutamine

For mitochondrial assays add:

* 0.5-10 mM Sodium pyruvate
* 1-4 mM L-Glutamine
* 2-5 mM Glucose

To make base media add the following:

{| 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
|-
| Magnesium sulfate
| 1 M
| 400 uL
| 0.8 mM
|-
| Calcium chloride
| 1 M
| 900 uL
| 1.8 mM
|-
| Sodium Chloride
| 4 M
| 17.8 mL
| 143 mM
|-
| Potassium chloride
| 1 M
| 2.7 mL
| 5.4 mM
|-
| Sodium phosphate
| 200 mM
| 2.28 mL
| 0.91 mM
|}

Add ~450 mL water, bring pH to 7.4 carefully (there is very little buffering capacity, so add NaOH slowly). Bring final volume up to 500 mL and sterile filter into an autoclaved bottle. Store at 4C.