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Simple Western Size

The Simple Western Size assay is an automated Western blot — no gels, no transfer devices, no blots, no film, and no manual analysis! Just load your samples in a Simple Western system and press start—it's a complete, walk-away solution for molecular weight-based protein separation.

 

What Types of Size-Based Assays Are Possible on Simple Western?

 

Simple Western size-based immunoassays and total protein assays are super straightforward. The initial sample preparation for all size-based assays is very similar to a traditional Western Blot where the sample is prepared in an SDS sample buffer that contains a reducing agent. Simple Western just requires additional fluorescent standard mix used to determine the molecular weight of your protein of interest. Samples are then heat-denatured and pipetted onto a Simple Western plate along with required detection reagents. From there, just load your capillary cartridge and sample plate and push run. That’s it! You can design your experiment to probe for your protein target(s) of interest in an immunoassay and/or use one of several total protein assay chemistries to visualize all the protein bands in your sample. Also view our RePlex™ options for second sequential immunoprobing round.

 
 
 
 



 


 
 
 
 
 
 
 
 
 
 
 
 
IR
Fluorescence
Detection
NIR
Fluorescence
Detection
HRP
Chemiluminescence
Detection
Green
Fluorescence
Detection
IR
Fluorescence
Detection
HRP
Chemiluminescence
Detection
NIR
Fluorescence
Detection
 
Green
Fluorescence
Detection
+

Multiplexed Western Blotting Assays

All size-based Simple Western immunoassays offer the flexibility to multiplex and detect multiple proteins per sample, giving you more data from the same sample. Plus, you can combine single or multiplexed immunoassays with total protein assays to normalize your immunoassay protein expression data with total protein expression on each sample. Just load your samples and off to the races you go!

Singleplex Assays

Multiplex Assays

Chemiluminescence
Immunoassay

or

Fluorescence
Immunoassay
or

Total Protein
Assay

Size based multiplexing

2+ immunoassays
in 1 detection channel

Detection channel
based multiplexing

Superplexing

Sequential
multiplexing

RePlexing

Chemiluminescence
immunoassay 1
+

Chemiluminescence
immunoassay 2

Fluorescence
immunoassay 1
+

Fluorescence
immunoassay 2

Chemiluminescence
Immunoassay(s)
+

Fluorescence
Immunoassay(s)
+

Total protein assay using the Protein Normalization module

Probe 1: Fluorescence/
Chemiluminescence
immunoassay(s)
+

Probe 2: Fluorescence/
Chemiluminescence
immunoassay(s)
or
Total protein assay

Advantages Of Increasing Assay Plex

Superplexing

All the advantages of multiplexing, plus:

  • Chemi, fluorescence, and total protein normalization (PN) in the same capillary

RePlexing

All the advantages of Superplexing, plus:

  • Two independent immunoprobes in the same capillary
  • Greater flexibility in antibody selection
  • Higher sensitivity protein normalization
  • Quantify phosphorylated target and total target

Multiplexing

  • More data points per sample
  • Multiple protein detection
  • Faster time to data
  • Lower cost / sample

Simple Western Multiplex Assay Compatibility Chart

See how Simple Western multiplex assays are compatible in the table below. You can also build your own assay using the Simple Western Kit Builder.

Assay Type

Instrument

Detection

Separation Module

Detection Modules

RePlex

Size-Based Multiplex Peggy Sue, Sally Sue • Chemi Separation Modules for Peggy Sue or Sally Sue • Chemi Detection Modules or Total Protein Detection Module (DM-TP01) No

Size-Based Multiplex

Jess, Abby

• Chemi

Std Separation Modules

Fluor Separation Modules

• Chemi Detection Modules or Total Protein Detection Module (DM-TP01)

Yes

Channel-Based Multiplex

Jess

• Chemi

• Fluor

Fluor Separation Modules

• Chemi Detection Modules or Total Protein Detection Module (DM-TP01)

• NIR / IR Detection Modules

Yes

Superplex

Jess

• Chemi

• Fluor

Fluor Separation Modules

• Protein Normalization Module (DM-PN02)

• Chemi Detection Modules

• NIR / IR Detection Modules

No

Stellar

Jess

• Chemi

• Fluor

Fluor Separation Modules

• Chemi Detection Modules or Stellar Total Protein Detection Module (DM-TP03)

• Stellar NIR / IR Detection Modules

No
  • Separation Modules for Peggy Sue and Sally Sue include all MW ranges (2-40, 12-230, 66-440 kDa)
  • Separation Modules for Jess and Abby include all MW ranges (2-40, 12-230, 66-440 kDa) and capillary cartridge formats (2 x and 8 x of 13 and 25 capillary cartridges)
  • NIR/IR Detection Modules include Anti-Mouse and Anti-Rabbit Modules
  • Chemi Detection Modules include Anti-Mouse, Anti-Rabbit, Anti-Goat, Anti-Human, Biotin Modules, and No-Secondary Detection Modules

Applications Specific Kit: SARS-CoV-2 Multi-Antigen Serology Module

The SARS-CoV-2 Multi-Antigen Serology Module is designed for simultaneous detection of human serum IgG reactivity with 5 viral antigens of interest for characterization of the COVID-19 immune response, including the nucleocapsid, the full-length spike protein, as well the spike protein S2, S1, and S1 RBD subunits. Analyze up to 24 samples within one 3-hour run to advance understanding of the human immune response to SARS-CoV-2! Learn more about the benefits of the SARS-CoV-2 serology assay.

Proof of concept studies have also shown that SARS-CoV-2 Multi-Antigen Serology Module for Jess/Abby/Wes can be used to monitor the COVID-19 immune response in animal systems such as mice when the appropriate secondary antibody is chosen. Learn more about the benefits of the SARS-CoV-2 serology assay.