HHBS (Hank's Buffer with Hepes) Preparation and Recipe

HHBS is a general purpose buffer that combines the stability of Hank's buffer with the good buffering capacity of HEPES buffering solution (one of Good's buffers). HHBS is used extensively for cell culture, but is sensitive to light.

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To prepare L of HHBS (Hank's Buffer with Hepes):
Change the value in the textbox above to scale the recipe volume

Table 1. Required components

Component	Amount	Concentration
Calcium Chloride (mw: 110.98 g/mol)	0.14 g	0.00126 M
Magnesium Chloride Hexahydrate (mw: 203.30 g/mol)	0.1 g	0.00049 M
Magesium Sulfate Heptahydrate (mw: 246.47 g/mol)	0.1 g	0.00041 M
Potassium Chloride (mw: 74.55 g/mol)	0.4 g	0.00533 M
Potassium Phosphate Monobasic (mw: 136.09 g/mol)	0.06 g	0.00044 M
Sodium bicarbonate (mw: 84.01 g/mol)	0.35 g	0.00417 M
Sodium chloride (mw: 58.44 g/mol)	8 g	0.138 M
Sodium Phosphate Dibasic (mw: 141.96 g/mol)	0.048 g	0.00034 M
D-Glucose (Dextrose) (mw: 180.16 g/mol)	1 g	0.00556 M
Hepes (mw: 238.30 g/mol)	4.76 g	0.02 M

Prepare 800 mL of distilled water in a suitable container.
Add 0.14 g of Calcium Chloride to the solution.
Add 0.1 g of Magnesium Chloride Hexahydrate to the solution.
Add 0.1 g of Magesium Sulfate Heptahydrate to the solution.
Add 0.4 g of Potassium Chloride to the solution.
Add 0.06 g of Potassium Phosphate Monobasic to the solution.
Add 0.35 g of Sodium bicarbonate to the solution.
Add 8 g of Sodium chloride to the solution.
Add 0.048 g of Sodium Phosphate Dibasic to the solution.
Add 1 g of D-Glucose (Dextrose) to the solution.
Add 4.76 g of Hepes to the solution.
Add distilled water until the volume is 1 L.

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

This online tool may be cited as follows

MLA	"Quest Calculate™ HHBS (Hank's Buffer with Hepes) Preparation and Recipe." AAT Bioquest, Inc., 21 Dec. 2024, https://www.aatbio.com/resources/buffer-preparations-and-recipes/hhbs-hanks-buffer-with-hepes.
APA	AAT Bioquest, Inc. (2024, December 21). Quest Calculate™ HHBS (Hank's Buffer with Hepes) Preparation and Recipe. AAT Bioquest. https://www.aatbio.com/resources/buffer-preparations-and-recipes/hhbs-hanks-buffer-with-hepes.
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This online tool has been cited in 1 publications, including

Identification of Autophagy as a Functional Target Suitable for the Pharmacological Treatment of Mitochondrial Membrane Protein-Associated Neurodegeneration (MPAN) In Vitro
Authors:
Journal: Pharmaceutics (2023): 267

HHBS (Hank's Buffer with Hepes) Preparation and Recipe

References

This online tool may be cited as follows

MLA

APA

This online tool has been cited in 1 publications, including