H-50 Buffer Preparation and Recipe

H-50 buffer is commonly used for washing and resuspending cells. It contains HEPES, one of the Good's zwitterionic buffers.

To prepare L of H-50 Buffer:
Change the value in the textbox above to scale the recipe volume

Table 1. Required components

Component	Amount	Concentration
Hepes (mw: 238.30 g/mol)	4.76 g	0.02 M
Potassium Chloride (mw: 74.55 g/mol)	3.73 g	0.05 M
Magnesium sulfate (mw: 120.37 g/mol)	0.12 g	0.001 M
Sodium chloride (mw: 58.44 g/mol)	0.58 g	0.01 M
Sodium bicarbonate (mw: 84.01 g/mol)	0.42 g	0.005 M
Sodium phoshpate monobasic dihydrate (mw: 156.01 g/mol)	0.156 g	0.001 M

Prepare 800 mL of distilled water in a suitable container.
Add 4.76 g of Hepes to the solution.
Add 3.73 g of Potassium Chloride to the solution.
Add 0.12 g of Magnesium sulfate to the solution.
Add 0.58 g of Sodium chloride to the solution.
Add 0.42 g of Sodium bicarbonate to the solution.
Add 0.156 g of Sodium phoshpate monobasic dihydrate to the solution.
Add distilled water until the volume is 1 L.
Adjust the pH to 7.0 with HCl or NaOH, as appropriate. Autoclave and store cold or frozen.

Physiological Buffer

pH Buffering

Sample Preparation

BioAssays

Misc

Cell/Culture/Growth Media

Gel Electrophoresis

Name ▲ ▼
Extracellular Saline (1x, pH 7.3)
HBSS (Hank's Balanced Salt Solution)
HEPES Buffer (1 M, 7.5 pH)
HHBS (Hank's Buffer with Hepes)
PBS (Phosphate Buffered Saline) (1X, pH 7.4)
PBS with Ca2+and Mg2+ (D-PBS)
Tyrode's Solution, Acidic

Name ▲ ▼
ACES Buffer (0.1 M, 6.7 pH)
Acetate Buffer (0.1 M, pH 5.0)
Acetate Buffer (pH 3.6 to 5.6)
ADA Buffer (0.5 M, 6.6 pH)
Ammonium Acetate
Ammonium Bicarbonate (50 mM, pH 7.8)
Ammonium Sulfate, Saturated
BES-Buffered Saline (2X) (0.05 M, pH 6.95)
Bicine (1 M, pH 8.26)
Bis-Tris Buffer (1 M, 6.5 pH)
Borate Buffer
CAPS (0.5 M, pH 10.4)
Carbonate-Bicarbonate Buffer (pH 9.2 to 10.6)
CHES (0.5 M, pH 9.5)
Citrate Buffer (0.1 M, pH 6.0)
Citrate Buffer (pH 3.0 to 6.2)
Citrate-Phosphate Buffer (0.15 M, pH 5.0)
Citrate-Phosphate Buffer (110 mM, pH 5.6)
Citrate-Phosphate Buffer (40 mM, pH 5.6)
Diethanolamine (1 M, pH 9.8)
EBSS (magnesium, calcium, phenol red) (pH 7.0)
Glycine (0.1 M, pH 2.2)
Glycine-HCl Buffer (0.1 M, pH 3.0)
Glycine-Sodium Hydroxide Buffer (0.08 M, pH 10)
HEPPSO (1 M, pH 7.85)
Hydrochloric Acid-Potassium Chloride Buffer (0.1 M, pH 2.0)
Imidazole-HCl Buffer (0.05 M, pH 7.0)
Maleic Acid (1 M, pH 7.5)
MES (0.5 M, pH 6)
MOPS Buffer (10X) (0.2 M, pH 7)
MOPSO Buffer (0.5 M, 6.9 pH)
Phosphate Buffer (pH 5.8 to 7.4)
PIPES Buffer (1 M, 6.8 pH)
Potassium Phosphate Buffer (1 M, pH 6.5)
Potassium Phosphate (pH 5.8 to 8.0)
Sodium Acetate (3 M, pH 5.2)
Sodium Borate Buffer (1 M, pH 8.5)
Sodium Phosphate Solution
SSC buffer (20X) (3 M, pH 7)
TAE (1 M, pH 8.6)
TB Buffer
TBS (1 M, pH 7.4)
TES Buffer (1 M, 7.5 pH)
Tricine (1 M, pH 8.05)
Tris Buffer (1 M, pH 7.2)
Tris-Buffered Saline (TBS, 0.1 M)
Tris-citrate Buffer (pH 8.0)
Trizma® Buffer (pH 7.0 to 9.2)

Name ▲ ▼
ACK Lysis Buffer
Alkaline Sodium Thiosulfate
BAC DNA Microinjection Buffer
DEAE-dextran
EDTA Solution
Elution Buffer
Gey's Balanced Salt Solution
H-50 Buffer
Lysis Solution
Osmium Tetroxide Solution
Paraformaldehyde Solution (8%)
TE Buffer 10X
Trichloroacetic Acid (TCA)

Name ▲ ▼
Anesthetic (MS222) Solution
Bradford Stock Solution
5-Fluorocytosine (5-FC) Solution
Chloroquine
Cresol Red Solution
Ethidium Bromide Solution
Immunofluorescence Quenching Solution
Mayer's Hemalum
Nuclear Fast Red Solution
Pyronin Y (PY) Solution
SSC (Northern Blot, 20x)
SSPE (Blot, 20x, pH 7.4)

Name ▲ ▼
Dithiothreitol (DTT) (1 M)
Sodium Iodide/Sodium Sulfite Solution

Name ▲ ▼
Bath Solution (Ca2+/Mg2+ Free)
Calcium Chloride Stock Solution
Carbenicillin Solution
Chelation Medium
Digestion Medium
Embryonic Medium
IPTG Solution
M9 Minimal Salts
Modified Barth's Saline (MBS)
Penicillin/Streptomycin/Chloramphenicol Antibiotic Mix
Yeast Two Hybrid (Y2H) Media, Amino Acid Dropout Mixes

Name ▲ ▼
Acrylamide Stock Solution
Electrode Buffer
Electrophoretic Transfer Buffer (20x)
Imidazole-SDS Solution
SDS-PAGE SDS Running Buffer (10x)
Sodium Carbonate Transfer Buffer (40x, pH 9.5)
Tris/EACA Buffer
Tris-Glycine Transfer Buffer (20x)

References

MLA	"Quest Calculate™ H-50 Buffer Preparation and Recipe." AAT Bioquest, Inc., 21 Dec. 2024, https://www.aatbio.com/resources/buffer-preparations-and-recipes/h-50-buffer.
APA	AAT Bioquest, Inc. (2024, December 21). Quest Calculate™ H-50 Buffer Preparation and Recipe. AAT Bioquest. https://www.aatbio.com/resources/buffer-preparations-and-recipes/h-50-buffer.
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