What makes the filtration process superior to standard 0.22 micron filtered media?

FluxMPS uses four sequential filters - 0.1 micron prefiltration I, 0.04 micron final filtration I, 0.1 micron prefiltration II, and 0.04 micron final filtration II (polish) - reaching a 0.04 micron final cut-off, five times finer than the 0.22 micron membranes used in conventional sterile filtration.

Does this medium require a CO2 incubator?

5 percent CO2 recommended. The 2000 mg/L sodium bicarbonate buffering system is calibrated for a 5 percent CO2 atmosphere; the added 25 mM HEPES provides supplementary buffering during brief periods outside the incubator.

FluxMPS RPMI 1640 Medium with 25mM HEPES: 1X Liquid

Q: Can DCP-RPMIH1X be supplemented with serum, growth factors, or other components?

Yes. FBS (5-10 percent), serum-free supplements, growth factors, or antibiotics can be added. Serum and other protein-containing supplements should be pre-filtered through a 0.2 micron low-protein-binding PES or PVDF membrane before addition - never through a 0.04 micron membrane, which will retain serum proteins and lipoproteins. Contact support@diagnocine.com for custom co-formulation.

Q: What endotoxin level should I expect, and how is it verified?

The release specification is less than 0.05 EU/mL, verified by LAL assay per USP 85 on every manufacturing batch (assay sensitivity 0.005 EU/mL). This is relevant for immune cell assays, where endotoxin can activate TLR4/NF-kappaB signaling and alter lymphocyte activation state independently of experimental conditions.

Q: Is a Certificate of Analysis (CoA) available, and what does it contain?

Yes. Full CoA per batch covers: appearance, pH (USP 791), osmolality (USP 785), sterility (USP 71), endotoxin (USP 85), mycoplasma filtration status, particulate count (USP 788 Method 1), and raw-material traceability. Request at support@diagnocine.com.

verified ISO 13485 Certified Manufacturing

FluxMPS™ RPMI 1640 Medium with 25mM HEPES

L-Glutamine Sodium Bicarbonate Phenol Red HEPES (25 mM) Calcium Magnesium Glucose (2 g/L) Sodium Pyruvate

FluxMPS™ DCP-RPMIH1X is a Microfluidics Suitable, quadruple-stage ultra-filtered (0.1 µm ×2 + 0.04 µm ×2) RPMI 1640 + 25 mM HEPES formulation engineered for hematopoietic and lymphocyte cell models on organ-on-a-chip (OoC) and microphysiological system (MPS) platforms. The train reaches a 0.04 µm final cut-off — five times finer than the 0.22 µm membranes used in conventional sterile filtration. Dual-buffered with 25 mM HEPES (pKa 7.3 at 37°C) and sodium bicarbonate, it maintains stable pH during both incubator culture and open-air handling. Formulation: [+] 2 g/L Glucose, [+] L-Glutamine, [+] Sodium Pyruvate, [+] Sodium Bicarbonate, [+] 25 mM HEPES.

Glucose 2000 mg/L (2.0 g/L) supports high-glycolytic hematopoietic and lymphocyte cell lines
25 mM HEPES (pKa 7.3 at 37°C) plus sodium bicarbonate — dual buffering stable outside CO₂ incubators
Glutathione (reduced), 1.000 mg/L — antioxidant support for redox-sensitive lymphocyte cultures
Quadruple-stage filtration train (0.1 µm ×2 + 0.04 µm ×2) reaching a 0.04 µm final cut-off
Endotoxin release specification < 0.05 EU/mL, LAL assay (USP <85>)
Manufactured under an ISO 13485:2016 quality management system; final QC at Diagnocine, Totowa, NJ
42 formulation components across inorganic salts, amino acids, vitamins and other constituents, released per lot
Custom pH, HEPES concentration, and nutrient modifications available on request

CAT. NO.

DCP-RPMIH1X | Cell Culture Media UNSPSC: 41116155 | Commodity: Molecular biology and cell culture growth media | (UNv260801)

RPMI 1640 + 25 mM HEPES — 1X Liquid

Media familyRPMI 1640 + 25 mM HEPES
Formulation[+] Glucose, [+] L-Glutamine, [+] Sodium Pyruvate, [+] Sodium Bicarbonate, [+] HEPES 25 mM
AppearanceRed-colored, clear solution
pH (USP <791>)7.4
Osmolality (USP <785>)280–320 mOsm/kg H₂O
Endotoxin (USP <85>)< 0.05 EU/mL
Filtration0.1 µm ×2 + 0.04 µm ×2 (Quadruple-stage)
Storage2–8°C, protect from light
Shelf Life12 months from date of manufacture, unopened
CO₂ Requirement5% CO₂ recommended

ISO 13485:2016 USP <85> <785> <788> RUO

Why FluxMPS™

Engineered where standard media fails

Conventional 0.22 µm–filtered RPMI passes mycoplasma-sized particles, subvisible particulates, and endotoxin fragments that activate TLR4 and confound immune cell assays. FluxMPS™ is built to control these failure modes at the filtration stage.

filter_alt

Microchannel-safe purity

0.04 µm final filtration; USP <788> Method 1 (light obscuration) particulate compliance. Particle-controlled media for suspension immune cell OoC and flow cytometry workflows.

science

Immune cell–optimized formulation

RPMI 1640 contains reduced glutathione (antioxidant) and a balanced profile of vitamins and amino acids suited to lymphocyte and hematopoietic cell culture.

water_drop

Ultrapure-grade water

Type 1 water (18.2 MΩ·cm) supports low trace-metal and organic-carbon background for sensitive lymphocyte culture.

visibility

Low background for imaging

Ultra-low particulate baseline supports confocal microscopy and biosensor-integrated OoC platforms. This formulation contains phenol red; a phenol red–free variant is available on request for autofluorescence-sensitive imaging.

shield

Endotoxin release specification

< 0.05 EU/mL by LAL assay (USP <85>) — relevant to controlling TLR4-mediated activation artefacts in lymphocyte and monocyte assays.

tune

Customization on demand

pH, nutrient concentrations, HEPES, and component modifications available. Contact support@diagnocine.com.

Purity Architecture

Quadruple-stage filtration system

Four serial filtration stages — two dedicated 0.1 µm prefilter + 0.04 µm final-filter pairs — reaching a final 0.04 µm cut-off, five times finer than the 0.22 µm membranes used in conventional sterile filtration.

1

0.1 µm Prefiltration I

Removes large particulate, cell debris and protein aggregates; protects the first 0.04 µm final-filter cartridge.
2

0.04 µm Final filtration I

First 0.04 µm pass; retains sub-micron particulates and microaggregates that pass a conventional 0.22 µm filter, including the 0.2–0.3 µm mycoplasma size range.
3

0.1 µm Prefiltration II

Second dedicated prefilter, protecting the second 0.04 µm cartridge for full-redundancy processing.
4

0.04 µm Final filtration II — Polish

Ultimate polishing filter prior to aseptic fill & finish.

Performance vs. conventional media

A quadruple-stage train (0.1 µm ×2 + 0.04 µm ×2) reaches a 0.04 µm final cut-off, five times finer than the 0.22 µm membranes used for conventional sterile filtration.

4

Sequential filtration passes (0.1 µm ×2 + 0.04 µm ×2)

0.04

µm final pore size — sub-mycoplasma-retentive polishing

Sterility & Mycoplasma: No growth after 14-day incubation (USP <71>). Mycoplasma risk is mitigated by 0.1 µm mycoplasma-retentive filtration (not tested per lot); mycoplasma organisms are typically 0.2–0.3 µm in diameter.

Grade: This product is Microfluidics Suitable, filtered to a 0.04 µm final cut-off. It is not an MPS Grade product — that designation is reserved for the 0.01 µm ultra nano-filtered line, which adds 0.02 µm and 0.01 µm stages after the 0.04 µm polish. For applications requiring the 0.01 µm cut-off, contact support@diagnocine.com.

FluxMPS DCP-RPMIH1X RPMI 1640 with 25mM HEPES Quadruple-stage filtration system 0.1 micron x2 plus 0.04 micron x2 for organ-on-a-chip and microfluidic immune cell applications Diagnocine — **Figure 1.** FluxMPS™ Quadruple-stage filtration system (0.1 µm ×2 + 0.04 µm ×2).
© Diagnocine® — DCP-RPMIH1X

Applications

Immune cell OoC and hematopoietic applications

FluxMPS™ DCP-RPMIH1X — RPMI 1640 + 25 mM HEPES — delivers 0.04 µm filtered, Microfluidics Suitable purity for hematopoietic and lymphocyte OoC applications.

Automated Bioreactors & Robotics

Next-Generation System Uptime

An optional 0.01 µm (10 nm) MPS Grade variant — adding 0.02 µm and 0.01 µm stages beyond this product's 0.04 µm polish — is available on request for automated bioreactors and robotics.

Total Particulate Exclusion: 10 nm filtration removes nanoparticulate aggregates
Valve & Sensor Protection: Reduces micro-fouling risk in automated perfusion systems
Extended Perfusion Stability: Consistent nutrient delivery over long-duration culture

Inquiry Required: Contact support@diagnocine.com for the 0.01 µm MPS Grade variant.

Immunology

T Cell & Lymphocyte Culture

RPMI 1640 is the standard for primary T cells, B cells, NK cells, and monocytes. FluxMPS™ 0.04 µm filtration reduces particulate load associated with TLR4 activation artefacts in immune assays.

T cellsB cellsNK cellsPBMC

Cancer Biology

Leukemia & Lymphoma Lines

RPMI supports NCI-60 cancer lines, Jurkat, Raji, K562, HL-60, and other hematopoietic cancer lines where a DMEM-based medium would alter proliferation and signaling.

JurkatRajiK562HL-60

Microfluidics

Immune Cell OoC

0.04 µm filtered RPMI for tumor-immune interaction chips, vascular-immune OoC, and lymph node-on-chip platforms where particulate control matters.

Tumor-immune chipLymph node OoCMPS

Immunotherapy

CAR-T & TIL Expansion

Low endotoxin release specification (< 0.05 EU/mL) is relevant to controlling LPS-driven T cell activation artefacts in CAR-T manufacturing and TIL expansion protocols.

CAR-TTILTCR-T

Metabolomics

Immune Cell Metabolic Flux

Defined RPMI base for glycolysis stress testing and ¹³C isotope tracing of lymphocyte activation states. Not compatible with Agilent Seahorse XF assays, which require bicarbonate-free, phenol red–free medium.

¹³C tracingGlycolysis

Live-Cell Imaging

Flow Cytometry & Confocal

Ultra-low particulate purity supports flow cytometry and confocal imaging workflows for immune cell populations. This formulation contains phenol red; request our phenol red–free variant for autofluorescence-sensitive imaging.

Flow cytometryConfocalELISA

Technical Specifications

Analytical release specifications

Every lot released against the full specification matrix. CoA: support@diagnocine.com.

Physical & Chemical Parameters

Parameter	Specification
Formulation	[+] Glucose (2 g/L), [+] L-Glutamine, [+] Sodium Pyruvate, [+] Sodium Bicarbonate, [+] HEPES (25 mM), [+] Phenol Red, [+] Calcium, [+] Magnesium
Appearance	Red-colored, clear solution
Glucose	2000 mg/L (2.0 g/L)
HEPES	25 mM (pKa 7.3 at 37°C)
pH USP <791>	7.4
Osmolality USP <785>	280–320 mOsm/kg H₂O
Total ingredients	42

Sterility, Purity & Safety

Parameter	Specification
Endotoxin USP <85> BET	< 0.05 EU/mL
Sterility USP <71>	No growth / 14 days
Mycoplasma	0.1 µm mycoplasma-retentive filtration (not tested per lot)
Particulate ≥10 µm USP <788> Method 1	NMT 25/mL
Particulate ≥25 µm USP <788> Method 1	NMT 3/mL
Water purity	Type 1, 18.2 MΩ·cm
Manufacturing std.	ISO 13485:2016
Fill environment	ISO Class 5 (Class 100)

Storage, Handling & Logistics

Parameter	Specification
Storage temperature	2–8°C, protect from light
Freeze-thaw	Do not freeze
Shelf life	12 months from date of manufacture, unopened
Shipping condition	Cold pack
CO₂ requirement	5% CO₂ recommended (bicarbonate-buffered base; 25 mM HEPES provides additional pH stability during open-air handling)

Raw Materials & Regulatory

Parameter	Specification
Raw material grade	Reagent / cell culture grade
Traceability	Full lot traceability per ISO 13485
Manufacturing QMS ISO	ISO 13485:2016 certified
UNSPSC	41116155 — Molecular biology and cell culture growth media (UNv260801)
Regulatory alignment	21 CFR Part 820 (QMSR) aligned
Production method	Micro-batch, per-lot QC release
Intended use	Research Use Only (RUO)

Formulation

Full composition (mg/L)

RPMI 1640 + 25 mM HEPES: 42 ingredients verified per lot with CAS numbers for full raw-material traceability. Contains reduced glutathione and a balanced profile of vitamins and amino acids optimized for lymphocyte and hematopoietic cell culture.

Component	CAS Number	mg/L
INORGANIC SALTS
Calcium nitrate tetrahydrate	13477-34-4	100.000
Magnesium sulfate anhydrous	7487-88-9	48.840
Potassium chloride	7447-40-7	400.000
Sodium bicarbonate	144-55-8	2000.000
Sodium chloride	7647-14-5	6000.000
Sodium phosphate dibasic anhydrous	7558-79-4	800.000

Component	CAS Number	mg/L
AMINO ACIDS
Glycine	56-40-6	10.000
L-Arginine hydrochloride	1119-34-2	241.000
L-Asparagine	70-47-3	50.000
L-Aspartic acid	56-84-8	20.000
L-Cystine dihydrochloride	30925-07-6	65.200
L-Glutamic acid	56-86-0	20.000
L-Glutamine	56-85-9	300.000
L-Histidine hydrochloride monohydrate	5934-29-2	20.960
L-Hydroxyproline	51-35-4	20.000
L-Isoleucine	73-32-5	50.000
L-Leucine	61-90-5	50.000
L-Lysine hydrochloride	657-27-2	40.000
L-Methionine	63-68-3	15.000
L-Phenylalanine	63-91-2	15.000
L-Proline	147-85-3	20.000
L-Serine	56-45-1	30.000
L-Threonine	72-19-5	20.000
L-Tryptophan	73-22-3	5.000
L-Tyrosine Disodium Salt	69847-45-6	28.830
L-Valine	72-18-4	20.000

Component	CAS Number	mg/L
VITAMINS
Choline chloride	67-48-1	3.000
D-Biotin	58-85-5	0.200
D-Ca-Pantothenate	137-08-6	0.250
Folic acid	59-30-3	1.000
Niacinamide	98-92-0	1.000
Pyridoxine hydrochloride	58-56-0	1.000
Riboflavin	83-88-5	0.200
Thiamine hydrochloride	67-03-8	1.000
Vitamin B12	68-19-9	0.005
p-Amino benzoic acid (PABA)	150-13-0	1.000
OTHERS
D-Glucose	50-99-7	2000.000
Glutathione reduced	70-18-8	1.000
HEPES	7365-45-9	5958.000
Phenol red sodium salt	34487-61-1	5.300
Sodium Pyruvate	113-24-6	110.000
i-Inositol	87-89-8	35.000

Custom formulation: Contact support@diagnocine.com for DCP-RPMIH1X modifications.

Quality Assurance

Manufacturing & compliance

Every FluxMPS™ product is manufactured and released under a multi-layer quality system.

verified

ISO 13485:2016 Quality Management

Manufactured under an ISO 13485:2016–certified quality management system. Final QC at Diagnocine R&D Center, Totowa, NJ, USA.

water_drop

Ultrapure Type 1 Water

18.2 MΩ·cm Type 1 water supports low trace-metal and organic-carbon background.

biotech

Quadruple-Stage Sterile Filtration

0.1 µm ×2 + 0.04 µm ×2 filtration train with ISO Class 5 aseptic fill & finish.

assignment

Micro-Batch Precision

Small-batch, per-lot tested — no blending; Certificate of Analysis issued for every batch.

Endotoxin — USP <85> BET

Batch-level LAL assay; release specification < 0.05 EU/mL (assay sensitivity 0.005 EU/mL).

Particulate — USP <788> Method 1

Light obscuration: NMT 25/mL (≥10 µm), NMT 3/mL (≥25 µm).

Osmolality — USP <785>

Target: 280–320 mOsm/kg H₂O.

Documentation & CoA

Full CoA with raw-material traceability available on request.

Batch-level quality control. Endotoxin is controlled per manufacturing batch rather than per unit. Every batch is tested before release and must meet the release specification:

Endotoxin — LAL assay, USP <85> Bacterial Endotoxins Test; assay sensitivity 0.005 EU/mL; release specification < 0.05 EU/mL
pH, osmolality, conductivity, appearance and clarity
Sterility

A Certificate of Analysis is available on request at support@diagnocine.com.

Product Comparison

How DCP-RPMIH1X compares

FluxMPS™ DCP-RPMIH1X vs. conventional 0.22 µm–filtered RPMI formulations.

Parameter	DCP-RPMIH1X (FluxMPS™)	Conventional RPMI 1640 + 25mM HEPES (0.22 µm filtered)	Standard DMEM/RPMI (0.22 µm filtered)
Grade	Microfluidics Suitable	Not applicable (standard grade)	Not applicable (standard grade)
RPMI 1640 + 25mM HEPES dual-buffered formulation	check_circle Yes	cancel No	cancel No
Final filtration pore size	0.04 µm	0.22 µm	0.22 µm
Number of filtration stages	4 (Quadruple)	1	1
Mycoplasma-retentive filtration (0.1 µm)	check_circle Yes	cancel No	cancel No
Endotoxin (release specification)	FluxMPS™ — < 0.05 EU/mL	Corning classical liquid media — < 0.25 EU/mL Sigma-Aldrich DMEM complete medium — ≤ 2 EU/mL Gibco classical DMEM — Not specified (recorded per lot)
USP <788> Method 1 particulate tested	check_circle Yes	cancel No	cancel No
Water quality	Type 1, 18.2 MΩ·cm	Purified water	Purified water
Manufacturing QMS	ISO 13485:2016	ISO 9001 or none	ISO 9001 or none
Microfluidic channel compatible	check_circle Microfluidics Suitable	cancel Risk of clogging	cancel Risk of clogging
Custom formulation available	check_circle Yes	cancel No	cancel No

Comparison figures from published supplier specifications, accessed 2026-09-02. Suppliers that publish no numeric endotoxin specification are shown as “Not specified”.

FAQ

Frequently asked questions

Common questions about FluxMPS™ DCP-RPMIH1X — RPMI 1640 + 25 mM HEPES.

Is DCP-RPMIH1X suitable for organ-on-a-chip (OoC) applications?expand_more

Yes. DCP-RPMIH1X is Microfluidics Suitable, processed through a quadruple-stage filtration system (0.1 µm ×2 + 0.04 µm ×2) reaching a 0.04 µm final cut-off. RPMI 1640 with this filtration profile is suited to immune cell OoC, tumor-immune interaction chips, and lymphocyte perfusion models.

What makes the filtration process superior to standard 0.22 µm filtered media?expand_more

FluxMPS™ uses four sequential filters — 0.1 µm prefiltration I, 0.04 µm final filtration I, 0.1 µm prefiltration II, and 0.04 µm final filtration II (polish) — reaching a 0.04 µm final cut-off, five times finer than the 0.22 µm membranes used in conventional sterile filtration.

Why does this formulation include both HEPES and sodium bicarbonate, and how does that affect CO₂ requirements?expand_more

DCP-RPMIH1X is dual-buffered: 2000 mg/L sodium bicarbonate provides the primary buffering system (5% CO₂ recommended for optimal pH), while 25 mM HEPES (pKa 7.3 at 37°C) adds stability during open-air handling — flow cytometry preparation, assay plate setup, and multi-well stimulation experiments — where CO₂ control is inconsistent.

Does this medium require a CO₂ incubator?expand_more

5% CO₂ recommended. The 2000 mg/L sodium bicarbonate buffering system is calibrated for a 5% CO₂ atmosphere; the added 25 mM HEPES provides supplementary buffering during brief periods outside the incubator.

Can DCP-RPMIH1X be supplemented with serum, growth factors, or other components?expand_more

Yes. FBS (5–10%), serum-free supplements, growth factors, or antibiotics can be added. Serum and other protein-containing supplements should be pre-filtered through a 0.2 µm low-protein-binding PES or PVDF membrane before addition — never through a 0.04 µm membrane, which will retain serum proteins and lipoproteins. Contact support@diagnocine.com for custom co-formulation.

What endotoxin level should I expect, and how is it verified?expand_more

The release specification is < 0.05 EU/mL, verified by LAL assay per USP <85> on every manufacturing batch (assay sensitivity 0.005 EU/mL). This is relevant for immune cell assays, where endotoxin can activate TLR4/NF-κB signaling and alter lymphocyte activation state independently of experimental conditions.

Is a Certificate of Analysis (CoA) available, and what does it contain?expand_more

Yes. Full CoA per batch covers: appearance, pH (USP <791>), osmolality (USP <785>), sterility (USP <71>), endotoxin (USP <85>), mycoplasma filtration status, particulate count (USP <788> Method 1), and raw-material traceability. Request at support@diagnocine.com.

Scientific References

Supporting literature

Key publications supporting RPMI 1640 + 25 mM HEPES in immune cell culture and OoC applications.

Moore GE, et al. Culture of normal human leukocytes. JAMA. 1967;199:519–524. doi:10.1083/jcb.1.3.273
Huh D, et al. Reconstituting organ-level lung functions on a chip. Science. 2010;328:1662–1668. doi:10.1126/science.1188302
Bhatia SN, Ingber DE. Microfluidic organs-on-chips. Nat Biotechnol. 2014;32:760–772. doi:10.1038/nbt.2989
Novak R, et al. Robotic fluidic coupling and interrogation of multiple vascularized organ chips. Nat Biomed Eng. 2020;4:407–420. doi:10.1038/s41551-019-0497-x
Jang KJ, et al. Human kidney proximal tubule-on-a-chip. Integr Biol. 2013;5:1119–1129. doi:10.1039/c3ib40049b
Schimek K, et al. Integrating biological vasculature into a multi-organ-chip microsystem. Lab Chip. 2013;13:3588–3598. doi:10.1039/c3lc50217a
Luni C, et al. High-efficiency cellular reprogramming with microfluidics. Nat Methods. 2016;13:446–452. doi:10.1038/nmeth.3832
Sung JH, et al. Microfabricated mammalian organ systems. Lab Chip. 2013;13:1201–1212. doi:10.1039/c3lc41017j

FluxMPS™ RPMI 1640 Medium with 25mM HEPES: 1X Liquid