Fluorescent Western blotting

Fluorescent Western standardization blot
Fluorescent Western standardization blot

Ready-to-image three-color fluorescent blot

  • Fluorescently labeled primary and secondary antibodies fluoresce in the red, green, and blue fluorescent channels (like Cy2, Cy3, and Cy5 equivalent dyes)
  • Convenient – an easy way to evaluate the performance of your fluorescence imaging system
  • Reliable – quality controlled, with lot-specific statistics provided for every blot
SpectraDye Antibody Labeling Kits
SpectraDye Antibody Labeling Kits

Make your own fluorescent antibody in one easy step.

  • Quick and easy – one-step labeling in 30 minutes
  • Versatile - Labeled antibodies are compatible with immunofluorescence microscopy, flow cytometry, Western blotting
  • Flexible - choose from 6 commonly used fluorescent dyes
  • Save time and money – no secondary or extra steps, once you label your primary
  • Save antibody – labeling reaction requires as little as 10 µg antibody

 

Kit includes:
Antibody Labeling Buffer
SpectraDye Dye Solution*
Quenching Solution
Neutralization Buffer

Sufficient reagents to label up to 1 mg of antibody
*Except kit K-11060-010

 

Spectradyes

WesternBright MCF
WesternBright MCF

Multi-color fluorescent Western blotting in one kit

Try multicolor fluorescent western blotting today. Our kit includes everything except primary antibodies.

  • Multiplexing – visualize two proteins simultaneously!
  • Sensitivity –10x greater sensitivity than Cy dyes; pictogram detection
  • Quality Reagents – includes AdvanBlock™-PF blocking and AdvanWash /AdvanWash-IR for washing
  • Fast results – 3.5 hours for entire protocol
  • Compatible with imaging systems that detect Cy3 and Cy5 (MCF) or near-IR imaging systems (MCF-IR)
  • Fluorescent Detection – choose between visible (APC and RPE) or near IR (IR700 and IR800) kits with SpectraDye™ secondary antibodies

 

Kit Includes:
Secondary antibodies
Blocking solution
Washing solution
Pre-cut low-autofluorescence PVDF membranes
Background quenching sheets

 

AdvanBlock-PF blocking solution
AdvanBlock-PF blocking solution

Non-protein blocking solution for fluorescent and chemiluminescent Western blotting

  • Low background - Protein-free block buffer for lower background with certain antibodies
  • Optimized - Stabilizes fluorescence of WesternBright MCF fluorescent secondary antibodies
  • Versatile - Compatible with fluorescent and chemiluminescent detection
AdvanStain Total Fluorescent Protein Staining Kits
AdvanStain Total Fluorescent Protein Staining Kits

Rapid fluorescent stain for total protein normalization

 

  • Convenient – simple protocol that takes less than 20 minutes, no de-staining required
  • Versatile – compatible with chemiluminescent and IR700/IR800 fluorescent Western blotting
  • Accurate – linear across a broad range from 1-50μg of lysate, ideal for total protein normalization

AdvanWash washing solution
AdvanWash washing solution

Washing solutions optimized for use with WesternBright systems

  • Reliable – quality controlled wash buffer for reproducible results
  • Versatile – compatible with chemiluminescent and fluorescent Western blots
  • Convenient – save time using pre-made buffers; no more weighing, mixing, or adjusting pH

 

AdvanBlock-Fluor blocking solution
AdvanBlock-Fluor blocking solution

Optimized block solution 
for the best fluorescent Western blot signals

  • Enhanced signal – enhances Western blotting signals
  • Decreased background – decreased non-specific bands and lowered background
  • Sensitivity – get better signals for low protein sample concentrations
  • Convenience – Pre-made buffer means you save preparation and optimization time

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LightSaver™ Fluorescence Enhancing Solution
LightSaver™ Fluorescence Enhancing Solution

LightSaver™ Fluorescence Enhancing Solution

  • Enhances – fluorescent signal
  • Compatible – with PVDF and Nitrocellulose membranes
  • Convenient – ready-to-use solution
  • Compatible – with visible and near IR fluorescent westerns
FLASHBlot-SD transfer buffer
FLASHBlot-SD transfer buffer

Rapid high efficiency semi-dry transfer buffer

  • Fast - high ionic strength formulation allows for protein transfer in 3 to 10 minutes when used with a compatible high current semi-dry system
  • Compatible - use your existing high current semi-dry transfer apparatus
  • Reproducible - consistent transfer across entire blot
  • Versatile - use nitrocellulose or PVDF membranes to achieve transfers with low background and high sensitivity with both chemiluminescent and fluorescent Western blots
FLASHBlot transfer buffer
FLASHBlot transfer buffer

Ready-to-use wet transfer buffer
Designed for faster, more efficient wet protein transfer with increased sensitivity

FLASHBlot is not your average wet transfer buffer.

  • Faster than traditional wet buffers: better results in 20 minutes
  • More sensitive detection for low abundance proteins
  • Better detection of post-translationally modified proteins
  • Better transfer of ALL proteins, including high molecular weight proteins
  • Conveniently suitable for any wet transfer set-up

“ We were pleasantly surprised to see that with FLASHBlot we observed the same transferring levels at 25 min than with the lengthier transfer protocol… including for the 300 kDa protein. This reagent will shorten considerably our experimental time. ”  - Assistant Professor, UCSF

 

SpectraDye Secondary Antibodies
SpectraDye Secondary Antibodies

Fluorescently labeled secondary antibodies

  • Fluorescent detection –secondary antibodies with a range of fluorescent conjugates
  • Performance – strong signal and no cross-reactivity
  • Convenience – each antibody demonstrated to provide optimal results with Advansta's Western blotting systems
  • Flexibility – choose from visible or near IR fluorescent dyes and target species best suited to your application (see product listings)

2nd AB excitation emission chart

 

Transfer membranes
Transfer membranes

Convenient nitrocellulose and PVDF membranes for Western blotting

  • Performance – quality controlled for background-free Western blots
  • Convenience – membranes available pre-cut and as rolls
  • High signal-to-noise ratio – low background signal for clearer results
  • Flexibility – choose the right membrane for your Western blotting application

 

Membrane Options:

WesternBright PVDF-FL: fluorescent detection
WesternBright PVDF-CL: chemiluminescent detection; best stability for stripping and re-probing chemiluminescent blots
PVDF membranes available as rolls or pre-cut (7x9 cm, 10x15 cm, or 13x18 cm)

WesternBright NC: all-purpose nitrocellulose membrane
Nitrocellulose membranes available as rolls or pre-cut (8x10 cm)