Our broad portfolio consists of multiplex panels that allow you to choose, within the panel, analytes that best meet your needs. On a separate tab you can choose the premixed cytokine format or a single plex kit.
Cell Signaling Kits & MAPmates™
Choose fixed kits that allow you to explore entire pathways or processes. Or design your own kits by choosing single plex MAPmates™, following the provided guidelines.
The following MAPmates™ should not be plexed together:
-MAPmates™ that require a different assay buffer
-Phospho-specific and total MAPmate™ pairs, e.g. total GSK3β and GSK3β (Ser 9)
-PanTyr and site-specific MAPmates™, e.g. Phospho-EGF Receptor and phospho-STAT1 (Tyr701)
-More than 1 phospho-MAPmate™ for a single target (Akt, STAT3)
-GAPDH and β-Tubulin cannot be plexed with kits or MAPmates™ containing panTyr
.
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Select A Species, Panel Type, Kit or Sample Type
To begin designing your MILLIPLEX® MAP kit select a species, a panel type or kit of interest.
Custom Premix Selecting "Custom Premix" option means that all of the beads you have chosen will be premixed in manufacturing before the kit is sent to you.
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96-Well Plate
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Add Additional Reagents (Buffer and Detection Kit is required for use with MAPmates)
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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
Space Saver Option Customers purchasing multiple kits may choose to save storage space by eliminating the kit packaging and receiving their multiplex assay components in plastic bags for more compact storage.
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Immobilon® Membranes, Sandwiches and Blotting Filter Paper
Immobilon® PVDF membranes are the ideal transfer membranes for protein blotting applications.More
Immobilon® PVDF membranes are the ideal transfer membranes for protein blotting applications. Less
Millipore offers four different Immobilon® PVDF (Polyvinylidene Difluoride) membranes, each optimized for different protein blotting applications. We also now offer blotting sandwiches that feature pre-cut sheets of membrane and blotting filter paper.
Immobilon® membranes provide a number of advantages compared to nitrocellulose. They won’t crack or curl, and they can be cut without fracturing. They also have low background, broad solvent compatibility, and superior staining capabilities. In addition, they can be reprobed multiple times.
Immobilon®-P Membrane
0.45 µm pore size
Recommended for most western blotting, especially proteins >20 kDa
Millipore's Rapid Immunodetection Protocol reduces detection times by up to 2 hours by eliminating membrane blocking and several wash steps with no loss of sensitivity or specificity
Immobilon®-E Membrane
The only PVDF membrane that wets out in water—no alcohol required
0.45 µm pore size ideal for most western blotting applications
Save time and reduce waste by eliminating the alcohol pre-wet step
Immobilon-PSQ Membrane
0.2 µm pore size and a large internal structure
Higher protein adsorption and sequencing yields than other
membranes
Recommended for blotting proteins <20 kDa
Prevents blow-through of low molecular weight proteins
Immobilon-FL Membrane
The first transfer membrane optimized for fluorescence applications
Extremely low background improves sensitivity of all fluorescence detection protocols
Compatible with all commonly used fluorescent probes at all excitation and emission wavelengths
Ideal for multiplexing and chemifluorescence applications
Immobilon® NOW Rolls & Dispenser
Convenience of cut sheets with the flexibility of rolls, offered for each Immobilon® PVDF membrane
Mini or midi size with a single cut
No wasted membrane
More compact package saves space in your lab
Blotting Sandwiches
When you need to process multiple blots, blotting sandwiches are a convenient, time-saving choice. Our sandwiches include sheets of Immobilon®-E or Immobilon®-P membrane interleaved with pre-cut sheets of chromatography-grade blotting filter paper.
Blotting Filter Paper
Chromatography-grade blotting filter paper pre-cut to the most popular western blotting sizes.
Performance
Rapid Immunodetection Protocol with Immobilon-P Membrane
Step
Standard Immunodetection
Rapid Immunodetection
1. Block the membrane
1 h
None
2. Incubate with primary antibody
1 h
1 h
3. Wash the membrane
3 x 10 min
3 x 5 min
4. Incubate with secondary antibody
1 h
30 min
5. Wash the membrane
3 x 10 min
3 x 5 min
6. Add substrate
5 min
5 min
Total time
4 h 5 min
2 h 5 min
Immobilon-PSQ Membrane Prevents Blow-through
Molecular weight standards (lanes 1, 3, 5, 7) and calf liver lysate (lanes 2, 4, 6, 8) were transferred to Immobilon-P or Immobilon-PSQ membranes. A sheet of Immobilon-PSQ was placed behind the primary membranes to capture proteins that passed through (lanes 5 and 6 behind Immobilon-P; lanes 7 and 8 behind Immobilon-PSQ).
Immobilon-FL Membrane is Optimized for Fluorescence
Actin-tubulin multiplex detection on Immobilon-FL membrane. Rabbit muscle actin (red) was detected using rabbit anti-actin 1oAB and QDot® 655 goat anti-rabbit 2oAB. Porcine brain tubulin (green) was detected using mouse anti-tubulin 1oAB and QDot 565 goat anti-mouse 2oAB. Sensitivities down to 7 ng were observed for both analytes. Data provided by Quantum Dot Corporation.
Immobilon® PVDF membranes are offered in three types, each optimized for a different protein blotting application. Convenient blotting sandwiches feature pre-cut sheets of membrane and blotting filter paper. Immobilon® PVDF membranes have high protein adsorption, so you won’t lose proteins during transfer or reprobing. The open pore structure makes it easy to access bound proteins and remove unbound probes. In addition, Immobilon® PVDF membranes optimized for fluorescent blots dramatically increase signal-to-noise ratios for high sensitivity in quantitative, multiplexing applications.
Features & Benefits
Won’t crack, curl or fracture when cut
Low background
Superior staining capabilities
Can be reprobed multiple times
Applications
Western Blotting, Dot Blotting, Protein Sequencing; Compatible with Radioactive, Chromogenic, Chemiluminescent, Fluorescent, and Chemifluorescent Detection
Specifications
Ordering Information
Documentation
References
Reference overview
Application
Mobile Phase Preparation for UHPLC: Membrane Filtration Method Affects System Performance and Leaching of Extractable Impurities Subodh Kulkarn(1), Jesmi George(2) and Vivek Joshi(2) (1) Millipore India Pvt. Ltd., Bioscience Division, 50A, 2nd Phase, Ring Road, Peenya, Bangalore, India 560058 (2) Millipore Corp., Bioscience Division, 17 Cherry Hill Drive, Danvers, MA 01923 LCGC
2010
UHPLC/UPLC® is a revolutionary chromatography technique that is gaining wide acceptance among researchers due to improved resolution, shorter chromatographic runs, and the capability for doing fast method development. The presence of sub-2 µm particles in UHPLC columns provide these benefits but also poses challenges in sample and mobile phase preparation. Particulate impurities in the sample or mobile phase can cause backpressure buildup in the UHPLC system, causing system failure. In fact, most UHPLC instrument vendors recommend filtration of mobile phase using 0.2 µm filters, but there is a lack of data showing the benefits of filtration. In this article we describe the filtration of mobile phases through syringe filters of varying pore size and membrane type, followed by analysis by UHPLC and mass spectrometry. Our results clearly indicate that filtration of mobile phase components using the optimal membrane filter will help protect UHPLC systems from particulate impurities that may clog and shut down the system, increase the sensitivity of detection, and improve the accuracy of quantitation.
Quantitation of Protein on gels and blots by infrared fluorescence of Coomassie blue and fast green Luo S., Wehr N.B., Levine R.L. Analytical Biochemistry:350 (2006):233-238
2006
Immunoblotting (Western)
Role of the Small Heat Shock Proteins in Regulating Vascular Smooth Muscle Tone McLemore E.C., Tessier D.J., Thresher J., Komalavilas P., Brophy C.M J. Am. Coll. Surg. 2005, Vol 201 (1):30-36
2005
Pre-B-cell colony-enhancing factor is a secreted cytokine-like protein from the human amniotic epithelium. Ognjanovic S, Ku TL, Bryant-Greenwood GD. Am J Obstet Gynecol. 2005 Jul;193(1):273-82
2005
Western Blotting
A high-affinity reversible protein stain for Western blots Antharavally B.S., Carter, B., Bell, P.A., Mallia K. Analytical Biochemistry 2004,Vol 329:276-280
2004
Biochemical analysis of GABA receptor subunits alpha 1, alpha 5, beta1 beta2 in the hippocampus of patients with Alzheimer's disease neurophathology Rissman, R.A., Mishizen-Eberz A.J. N.,Wolfe, C.B.B., DeBlas A.L., Miralles C.P., Ikonomovic M.D., armstrong D.M. Neuroscience 120 (2003) 295-705
2003
Western Blotting
Proteomics reveals protein profile changes in doxorubicin treated MCF7 human breast cancer cells Cheng S.T., Pan T, L., Tsai Y.Ch, Huang C. M. Cancer letters 2002. vol 181:95-107
2002
Towards proteome-wide production of monoclonal antibody by phage display. Bin Liu, Lan Huang, Carina Sihlbom, Al Burlingame and James D. Marks. J Mol Biol. 2002 Feb 1;315(5):1063-73
2002
Mass Spectrometry Sample Prep
Characterization of retinoic acid receptor-deficient keratinocytes. Goyette Philippe; Chen Chang Feng; Wang Wei; Seguin Francois; Lohnes David(a) Journal of Biological Chemistry v 275 pg 16497-16505 June 2, 2000
2000
Protection of renal inner medullary epithelial cells from apoptosis by hypertonic stress-induced p53 activation Dmitrieva Natalia(a); Kultz Dietmar; Michea Luis; Ferraris Joan; Burg Maurice ; Journal of Biological Chemistry v 275, pg 18243-18247 June 16, 2000 Journal of Biological Chemistry v 275, pg 18243-18247 June 16, 2000
2000
Should I prewet MultiScreen Immobilon-P plates before use?
Yes. Use 15 ul of 70% ethanol or methanol to prewet the Immobilon P membrane. This membrane is hydrophobic and requires a prewet to allow liquid to pass through the membrane.
What is the protein binding capacity of Immobilon-P?
The protein (BSA) binding capacity for the Immobilon P membrane is 131 micrograms per sq. cm. of membrane.
What is the thickness of the Immobilon PSQ?
Immobilon PSQ is 200 microns thick.
Can ethanol be substituted for methanol in the Western Blot procedure?
Yes, it is acceptable to replace the methanol with ethanol as long as you substitue it 1 for 1. So 20% methanol would be replaced with 20% ethanol.
What is the smallest size of a protein or peptide which binds to the Immobilon-PSQ?
For the PSQ we do not suggest that they go lower than 2 kDa for the protein size. To help promote the transfer of the smaller proteins the methanol concentration of the transfer buffer can be increased to 20% (w/v) and the SDS lowered to 0.01%. The strength of the electric field can also be reduce by 50% to increase the proteins contact time with the membrane. Standard stains can be used such as Coomasie Blue and Ponceau. Keep in mind that some of the stain such as Coomasie Blue are not reversible.
How many times can I strip and reprobe Immobilon-P?
While 2-3 times is probably the limit, it is difficult to give an absolute number in regards to stripping and reprobing. This is because there are many factors to consider when it comes to protein binding to PVDF and primary antibody affinity to these proteins. Different proteins will have varying degrees of affinity to PVDF. This is based on factors discussed in our Protein Blotting Handbook (see TP001; Protein Binding section). One round of stripping may remove one protein and leave another intact. The same could occur when using different primary antibodies. Different antibodies will have varying affinities to different proteins. If carrying out several rounds of stripping and reprobing, one strategy might be to detect the least abundant proteins earlier leaving the higher abundant proteins later for detection.
What is the difference between Immobilon–FL and Immobilon-P?
Both membranes are made from PVDF. The difference in background fluorescence is due to proprietary modifications in the membrane manufacturing process.
How does the protein binding capacity and protein retention of Immobilon-FL compare to Immobilon-P?
Immobilon-FL’s protein binding capacity and protein retention are comparable to Immobilon-P.
Is Immobilon-FL better than Nitrocellulose membrane for Fluorescence detection?
Yes. Background fluorescence of Immobilon-FL is typically 2-5X lower than that of nitrocellulose, thus improving signal-to-noise ratio (sensitivity). Nitrocellulose membranes have other disadvantages. If allowed to dry out, they become brittle, tend to fracture and are difficult to handle. They are not recommended for stripping and re-probing. Nitrocellulose blots need to be scanned as soon as possible after detection, as diffusion of signal on a wet membrane may also occur.
What fluorescence-based detection methods can be used with Immobilon-FL?
Immobilon-FL can be used in Western blotting applications using either fluorescent dye-conjugated antibodies or chemi-fluorescence substrates. Western blots can be imaged in the visible or IR range. Single color detection or multiplexing for co-localization studies can be performed efficiently on Immobilon-FL Fluorescent proteins, e.g. green fluorescent protein, (GFP) or proteins tagged with GFP, blotted onto the membrane can be readily detected as well.
