Bio-Plex Pro™ Human Diabetes Assays

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Overview

Replacement Assays for Obsolete Catalog Numbers

The Bio-Plex Pro Human Diabetes assays are still based on magnetic beads, but they now offer expanded assay working ranges and shorter sample incubation times.

Please refer to the table below for a replacement item.

Obsolete Catalog Numbers	Comparable Replacement Catalog Numbers
Complete Kits Assay kit + Reagent kit + Diluent kit	Bio-Plex Pro Human Diabetes 10-plex Assay + Adiponectin and Adipsin
171-A4S01M 12- + 2-plex panel, 1 x 96-well	No direct equivalent; replaced by: 171-A7001M 10-plex assay, 1 x 96-well and 171-A7002M Adipsin and Adiponectin assays, 1 x 96-well
171-A4C01M 12-plex panel, 10 x 96-well	171-A7010M 10-plex assay, 10 x 96-well
171-B4C03M Adiponectin singleplex assay, 1 x 96-well 171-B4C05M Adipsin singleplex assay, 1 x 96-well	171-A7002M Adipsin and Asiponectin assays, 1 x 96-well 171-A7003M Adiponectin assay, 1 x 96-well 171-A7004M Adipsin assay, 1 x 96-well
Assay Kits Coupled beads + Detection antibodies + Standards	Bio-Plex Pro Human Diabetes 10-plex Assay + Adiponectin and Adipsin
171-A4001M 12-plex, 1 x 96-well 171-A4002M 12-plex, 10 x 96-well	No direct equivalent; replaced by singleplex sets (beads and detection antibodies) for the individual analytes. Requires reagent kit and standards.
171-B4003M Adiponectin singleplex assay, 1 x 96-well 171-B4004M Adiponectin singleplex assay, 10 x 96-well 171-B4005M Adipsin singleplex assay, 1 x 96-well 171-B4006M Adipsin singleplex assay, 10 x 96-well	171-A7002M Adipsin and Asiponectin assays, 1 x 96-well 171-A7003M Adiponectin assay, 1 x 96-well 171-A7004M Adipsin assay, 1 x 96-well
Reagent Kits Reagents + Instruction manual	Bio-Plex Pro Human Diabetes 10-plex Assay + Adiponectin and Adipsin Common to cytokine and diabetes assays
171-304030 1 x 96-well	171-304070 1 x 96-well
171-304031 10 x 96-well	171-304071 10 x 96-well
Diluent Kit 70 ml serum-based + 25 ml serum-free diluents	70 ml serum-based diluent For Adiponectin and Adipsin assays only
171-305040 1 x 96-well	171-305042 1 x 96-well
Standards Not available as stand-alone	(Adiponectin, Adipsin C-Peptide, Ghrelin, GIP, GLP-1, Glucagon, Insulin, Leptin, PAI-1, Resistin, Visfatin)
	171-D70001 1 pack of 12 analytes 171-D70050 50 pack of 12 analytes

New Bio-Plex Pro™ Human Diabetes Assays

Enhanced Performance —

Get your results faster with a shorter, one-hour incubation time — no need to incubate for two hours!

The assay working ranges have been expanded so more of your samples fall in range. See for yourself how normal and Type II diabetic samples are captured within the optimized assay range of Ghrelin.

Ghrelin Standard Curve and Analytes

Assay performed on the new Bio-Plex Manager software, 6.0. Click here to learn more about the new software.

Multiplexing — Combine with our Bio-Plex Pro cytokine assays

The cytokine analytes, IL-6 and TNF-α have been removed from the diabetes panel to allow multiplexing of diabetes with these and other cytokine assays. This is especially beneficial in the research areas of angiogenesis, immunology, and metabolic diseases.

Analytes Available

Obsolete	Current
Adiponectin	Adiponectin
Adipsin	Adipsin
C-Peptide	C-Peptide
Ghrelin	Ghrelin
GIP	GIP
GLP-1	GLP-1
Glucagon	Glucagon
IL-6
Insulin	Insulin
Leptin	Leptin
PAI-1	PAI-1
Resistin	Resistin
TNF-α
Visfatin	Visfatin

Convenience — New All-in-One kit!

Order one catalog number, and get one box with everything you need to start your assay:

Magnetic beads coupled with capture antibodies
Detection Antibody
Streptavidin-PE
Standards
Diluents & Buffers

Flexibility — You choose the analytes of interest:

Full diabetes panel
Singleplex assays of just the diabetes analytes of interest
Multiplex diabetes and cytokine assays

At-a-Glance Difference Between Obsolete and Current Diabetes Kits

	Obsolete	Current
Number of analytes	14	12 (IL-6 and TNF-α removed; all others remain the same)
Multiplex with cytokine assays	no	yes
Singleplex (single set) assays available	no	yes
Incubation time	2hr	1hr
All-in-one kit	no	yes

A new manual includes a Plate Layout Template and Calculation Worksheet

This manual is specific to current assays (ie: new bead regions and shorter incubation time) and should not be used with the obsolete diabetes kit.

Available Human Diabetes Assays

Bio-Plex Pro™ Human Diabetes Assays

Assay Menu*	10-Plex Panel (171-A7001M)	2-Plex Panel (171-A7002M)	Adiponectin All-in-One Format (171-A7003M)	Adipsin All-in-One Format (171-A7004M)	Bead Regions

Adiponectin **		•	•		64
Adipsin **		•		•	35
C-Peptide	•				72
Ghrelin	•				26
GIP	•				14
GLP-1	•				27
Glucagon	•				15
Insulin	•				12
Leptin	•				78
PAI-1	•				61
Resistin	•				65
Visfatin	•				22

* With the exception of adiponectin and adipsin, all assays may be ordered as either customized Express assays or as singleplex sets (vial of beads coupled to capture antibodies and vial of detection antibodies). Requires standard and Bio-Plex Pro reagent kit to perform an assay. To learn more about Express assays, go to www.bio-rad.com/bio-plex/assaybuilder.

** Due to different dilution scheme, adiponectin and adipsin are offered in an all-in-one format only.

Calculation Worksheet

Calculation Worksheet for Bio-Plex Pro Assays

Click here to access the worksheet

Calculation Worksheet for Bio-Plex Pro Assays

Assay Validation

Bio-Plex Pro Human Diabetes Assay Validation

Levels of biomarkers in sera of normal (♦) and type II diabetic () groups. Student’s t-test was used to determine statistical significance between groups. Black lines denote mean values. P values are indicated for markers that were significantly different from normal samples (P<0.05).

Linearity of Dilution

Linearity of dilution. Linearity of analyte measurements in either serum or plasma (3-fold) were measured using linear regression analysis. Result is shown for the glucagon assay diluted in human plasma. R² value was 0.98 or higher within the assay working range for all 12 assays in both serum and plasma.

Assay Working Ranges

Assay working ranges span the measured concentration ranges of a typical set of unknown samples. Data were analyzed using Bio-Plex Manager™ software version 6.0, using the standard curve optimization function. Recovery range specification was set to 80–120%. , standard points; Δ, type II diabetes samples; Δ, normal samples. Result is shown for ghrelin.

Specifications

Specifications for the NEW Bio-Plex™ Human Diabetes Assays

Incubation time	1 hr
Sample volume	12.5 µl
Accuracy (recovery range)	80–120%
Precision Intra-assay %CV Inter-assay %CV	≤10% ≤15%
Cross-reactivity	Negligible

Representative assay working ranges, assay sensitivity, and precision. The LLOQ, ULOQ, LOD, and inter-assay precision %CV are mean data determined from three independent multiplex assays in serum-based matrix. Intra-assay %CV is derived from one representative assay. LLOQ and ULOQ are defined as the boundary standard curve points in which the performance specifications of individual standard points were met for 10% intra-assay CV, 15% inter-assay CV, and recovery range of 80–120%. Exception is noted for GLP-1 with one point exhibiting 12% intra-assay CV; one standard point of Leptin exhibited 17% inter-assay CV. Data were generated using the magnetic workflow with the Bio-Plex Pro II wash station.


	Assay Working Ranges, pg/ml		Assay Sensitivity, pg/ml	Assay Precision
Targets	LLOQ	ULOQ	LOD	Intra-assay %CV	Inter-assay %CV

2-Plex Assays
Adiponectin	160.0	218,485	32.7	4	2
Adipsin	43.0	14,513	17.0	6	4

10-Plex Assays
C-Peptide	22.4	10,031	14.5	5	4
Ghrelin	16.6	8,502	1.2	4	2
GIP	11.2	22,895	0.8	3	4
GLP-1	31.3	16,000	5.3	6	3
Glucagon	15.7	3,500	4.9	5	6
Insulin	1.7	3,541	1.0	3	5
Leptin	11.5	129,107	3.1	3	4
PAI-1	8.8	47,850	2.2	5	4
Resistin	2.3	4,739	1.3	3	4
Visfatin	51.3	280,266	37.1	4	3

Assay Performance Definitions

Assay working range — the range of concentrations within which the assay is precise and accurate. Boundaries of the assay working range are the lower limit of quantitation (LLOQ) and the upper limit of quantitation (ULOQ). Precision is expressed as the coefficient of variation (%CV) at concentrations within the assay working range. Accuracy is expressed as a ratio of the observed concentration vs. the expected concentration of a known amount of spiked analyte within the assay working range.

Precision — the coefficient of variation (%CV) at concentrations within the assay working range

Accuracy — ratio of the observed concentration versus the expected concentration of a known amount of spiked analyte within the assay working range

Sensitivity (Limit of Detection, LOD) — the concentration of analyte for which the fluorescence intensity signal is 2-fold of the standard deviation above the background signal.

The Bio-Plex Pro™ human diabetes immunoassays offer reliable performance and a convenient format for studying human diabetes and obesity markers.

These assays are magnetic bead–based assays, available as either premixed panels or singleplex sets, that rapidly (three hr) detect multiple diabetes and obesity biomarkers in a single experiment, using as little as 12.5 µl of sample. The 10-plex assay detects:

C-peptide
Ghrelin
GIP
GLP-1
Glucagon
Insulin
Leptin
PAI-1 (total)
Resistin
Visfatin

Additionally, adiponectin and adipsin can also be assayed; they are available as either singleplexes or a 2-plex. Due to different dilution schemes, these biomarkers should be assayed separately from the ten markers in the panel.

Based on 6.5 µm magnetic beads, these assays are compatible with both vacuum and magnetic wash stations. Magnetic washing simplifies plate processing, provides increased throughput, and results in decreased variability and increased reproducibility.

Key Features

Magnetic beads for simplified plate processing
Results obtained in 3 hr due to short 1 hr incubation time
All-in-one, space-saving complete kit
Premixed standards for generation of curves over a broad dynamic range for each target

Other Assay Formats Available

Custom assays (mixed at the bench): available as Bio-Plex^® Express assays
Singleplex sets: each set includes a vial of coupled magnetic beads and a vial of detection bodies; requires a reagent kit (#171-304070) and a vial of standards (#171-D70001) for a complete assay

For multiplexing capabilities with cytokine assays, refer to bulletin 5975.

Instrument Compatibility

Bio-Plex 200 system
Bio-Plex 3D system
Bio-Plex Pro wash station
Bio-Plex Pro II wash station

Immunoassay for	Diabetes
Species	Human
Bead type	Magnetic
Custom assay options	Express
Incubation time, hr	1
Sample volume, µl	12.5
Accuracy (recovery range)	80–120%
Cross-reactivity	Negligible

Bio-Plex Pro Human Diabetes 10-Plex Assay

171-A7001M
1 x 96-well, includes conjugated magnetic beads, detection antibodies, standards, buffers, diluents, streptavidin-PE, filter plate, flat bottom plate, sealing tape, for the detection of 10 human diabetes biomarkers

List Price:		$1,950.00
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Bio-Plex Pro Human Diabetes 10-Plex Assay

171-A7010M
10 x 96-well, includes conjugated magnetic beads, detection antibodies, standards, buffers, diluents, streptavidin-PE, filter plate, sealing tape, for the detection of 10 human diabetes biomarkers

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Bio-Plex Pro Human Diabetes Adipsin and Adiponectin Assays

171-A7002M
1 x 96-well, includes conjugated magnetic beads, detection antibodies, standards, buffers, diluents, streptavidin-PE, filter plate, flat bottom plate, sealing tape, for the detection of human adiponectin and adipsin

List Price:		$810.00
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Bio-Plex Pro Human Diabetes Adiponectin Assay

171-A7003M
1 x 96-well, includes conjugated magnetic beads, detection antibodies, standards, buffers, diluents, streptavidin-PE, filter plate, flat bottom plate, sealing tape, for the detection of human adiponectin

List Price:		$420.00
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Bio-Plex Pro Human Diabetes Adipsin Assay

171-A7004M
1 x 96-well, includes conjugated magnetic beads, detection antibodies, standards, buffers, diluents, streptavidin-PE, filter plate, flat bottom plate, sealing tape, for the detection of human adipsin

List Price:		$425.00
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Bio-Plex Pro Human Diabetes C-Peptide Set

171-B7003M
1 x 96-well, includes conjugated magnetic beads and detection antibodies for the detection of human C-peptide, requires diabetes reagent kit and standards, which are not included

List Price:		$245.00
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Bio-Plex Pro Human Diabetes Ghrelin Set

171-B7004M
1 x 96-well, includes conjugated magnetic beads and detection antibodies for the detection of human ghrelin, requires diabetes reagent kit and standards, which are not included

List Price:		$245.00
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Bio-Plex Pro Human Diabetes GIP Set

171-B7005M
1 x 96-well, includes conjugated magnetic beads and detection antibodies for the detection of human GIP, requires diabetes reagent kit and standards, which are not included

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Bio-Plex Pro Human Diabetes GLP-1 Set

171-B7006M
1 x 96-well, includes conjugated magnetic beads and detection antibodies for the detection of human GLP-1, requires diabetes reagent kit and standards, which are not included

List Price:		$245.00
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Bio-Plex Pro Human Diabetes Glucagon Set

171-B7007M
1 x 96-well, includes conjugated magnetic beads and detection antibodies for the detection of human glucagon, requires diabetes reagent kit and standards, which are not included

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Bio-Plex Pro Human Diabetes Insulin Set

171-B7008M
1 x 96-well, includes conjugated magnetic beads and detection antibodies for the detection of human insulin, requires diabetes reagent kit and standards, which are not included

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Bio-Plex Pro Human Diabetes Leptin Set

171-B7009M
1 x 96-well, includes conjugated magnetic beads and detection antibodies for the detection of human leptin, requires diabetes reagent kit and standards, which are not included

List Price:		$245.00
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Bio-Plex Pro Human Diabetes PAI-1 Set

171-B7010M
1 x 96-well, includes conjugated magnetic beads and detection antibodies for the detection of human PAI-1, requires diabetes reagent kit and standards, which are not included

List Price:		$245.00
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Bio-Plex Pro Human Diabetes Resistin Set

171-B7011M
1 x 96-well, includes conjugated magnetic beads and detection antibodies for the detection of human resistin, requires diabetes reagent kit and standards, which are not included

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Bio-Plex Pro Human Diabetes Visfatin Set

171-B7012M
1 x 96-well, includes conjugated magnetic beads and detection antibodies for the detection of human visfatin, requires diabetes reagent kit and standards, which are not included

List Price:		$245.00
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Number	Description	Options
5405	Bio-Plex Multiplex System Brochure, Rev D	Click to download	[ Add to Cart ] Check Out
5507	Bio-Plex Multiplex System Ordering Information Sheet, Interactive PDF, Rev K	Click to download
5975	Multiplexing Compatibility of the Bio-Plex Pro Diabetes and Cytokine Assays, Rev A	Click to download	[ Add to Cart ] Check Out
5985	Development and Validation of Multiplex Assays for Human Diabetes Biomarkers, Rev A	Click to download
10010747	Instruction Manual, Bio-Plex Pro Diabetes Assays, Rev D	Click to download
10010747_45-49	Calculation Worksheet for Bio-Plex Pro Assays	Click to download
BR125FS	Second Generation Multiplex Immunoassays	Click to download
6335	Bio-Plex Multiplex Immunoassays Analyte Guide, Interactive PDF, Rev B	Click to download
6342	Bio-Plex Pro Diabetes Assay Panels Product Information Sheet, Rev A	Click to download
10024973	Bio-Plex Pro Assays Quick Guide 2, Rev A	Click to download

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