Overview

Krebs-Henseleit Buffer, 10x Stock - Krebs-Henseleit Buffer is the classical physiological perfusate for isolated mammalian organ/organoid studies. The GMExpression 10x carbonate-free stock is designed for shelf stability: bicarbonate is added by the user immediately before carbogen equilibration.

This avoids the calcium-carbonate precipitation problem that limits storage of complete bicarbonate-containing KHB while still giving the correct pH and oxygenation at the bench.

We also have

Tyrode's Solution · Artificial Cerebrospinal Fluid · GMExpression Buffer catalogue

Use and Applications

  • Organoid perfusion / manuplation
  • Langendorff heart perfusion and cardiovascular safety pharmacology
  • Isolated kidney, liver, intestine, and tissue-slice perfusion
  • Ex vivo pharmacology and organ bath studies
  • Physiological Perfusate Family bundle with aCSF and Tyrode's Solution

→Buffer Indicator Navigator

Use the GMExpression Buffer Indicator Navigator to find suitable pH ranges that match this buffer system, or start from a specific buffer and search for suitable working pH ranges for the intended experiment.

Package Contents

ItemSpecification
Bottle500 mL sterile bottle, makes 5 L of 1x working solution
Companion1 M NaHCO3, 100 mL, for point-of-use addition
DesignCarbonate-free stock for shelf stability
Sterility0.1 μm (mycoplasma-reduction grade) sterile-filtered; released mycoplasma-test-negative (PCR)

Composition - draft formulation / release specification

ComponentAmount / concentrationFunction
NaCl1.18 M in 10x stockPrimary ionic strength
KCl47 mM in 10x stockPhysiological potassium
CaCl2.2H2O25.2 mM in 10x stockPhysiological calcium
KH2PO412 mM in 10x stockPhosphate component
MgSO4.7H2O12 mM in 10x stockPhysiological magnesium
D-glucose110 mM in 10x stockMetabolic substrate
NaHCO3Added separately to 25 mM at 1xBicarbonate buffering after carbogen equilibration

Preparation

1Dilute 100 mL of 10x stock to 1 L with sterile ultrapure water (0.1μm filterated MQ water).
2Add 2.10 g NaHCO3 or 25 mL of 1 M NaHCO3 companion solution.
3Equilibrate with carbogen gas (95% O2 / 5% CO2) for approximately 30 minutes.
40.1/0.08 μm Filter the final 1x perfusate according to the organ-perfusion protocol and use within 24 hours.

Note

  • The 10x stock is expected to be acidic before bicarbonate addition. Do not adjust the stock itself to pH 7.4.

Cautions

Caution. Adding bicarbonate to concentrated stock can cause carbonate precipitation. Always dilute first, then add bicarbonate.
Caution. Perfusion workflows may require additional particulate filtration to prevent micro-embolism.

Sterile Filtration - Mycoplasma-Reduction Grade / Nanofiltration

Because this solution is used with living mammalian cells and tissue, it is sterilised through a 0.1 μm membrane rather than the conventional 0.22 μm sterilising grade. Mycoplasma are deformable enough to pass through 0.22 μm pores; where a 0.1 μm filteration gives a substantially lower mycoplasma margin, and each lot is released mycoplasma-test-negative by PCR.

If your experimental subjects are virus-sensitive cells, organoid or tissue samples, or if you wish to ensure that the experimental results are free from viral interference, we also provide Nanofiltration 1X stock (0.015 μm). Quotation separately via support@gmexpression.com；This is independent of the point-of-use particulate filtration (typically 0.2 μm) applied to the carbonated 1x perfusate to prevent micro-embolism.

Storage and Expiry · Safety

  • Store at 2-8 deg C away from direct light.
  • Unopened shelf life: 18 months from manufacture after QC validation.
  • Transport. Ships at ambient temperature in insulated packaging; brief transit excursions are tolerated. Refrigerate at 2-8 deg C on receipt and protect from freezing.
  • Handling and PPE. Wear a laboratory coat, nitrile gloves, and safety glasses for routine handling. The salt stock presents no significant chemical hazard; maintain aseptic technique. Consult the Safety Data Sheet before use.

References

  1. Krebs and Henseleit (1932), Hoppe-Seyler's Zeitschrift fuer Physiologische Chemie 210:33-66.
  2. Safety Data Sheet: SDS draft to be finalised before commercial release.

Frequently Asked Questions

Q1. Why carbonate-free?
Shelf-stable complete KHB is difficult because Ca2+ and carbonate can precipitate during storage. Point-of-use bicarbonate solves this.
Q2. Why use carbogen?
Carbogen sets the bicarbonate/CO2 equilibrium and oxygenates the perfusate for physiological use.
Internal review note. Product page draft prepared for GMExpression review. Final specifications, SDS, CoA fields, QC release limits, product photography, OpenCart options, and commercial pricing should be confirmed before publication.