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Thermo Scientific™ SuperSignal™ Western Blot Substrate Bundle, Femto + trial-size Atto

Catalog No. PIA45916
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100 mL + 20 mL bundle
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PIA45916 100 mL + 20 mL bundle
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Catalog No. PIA45916 Supplier Thermo Scientific™ Supplier No. A45916
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Description

Contains SuperSignal™ West Femto Maximum Sensitivity Chemiluminescent Substrate plus a sample size of SuperSignal™ West Atto Ultimate Sensitivity Chemiluminescent Substrate to enable trial and comparison of the newest and most sensitive substrate for low abundant targets.

  • SuperSignal West Femto
    • Sensitivity: detect low-femtogram amounts of protein
  • SuperSignal West Atto
    • Sensitivity: low-femtogram to high-attogram
    • Best for low abundant targets or precious samples

Specifications

Product Type Substrate
Detection Method Chemiluminescence
Form Liquid
For Use With (Application) Western Blot
Membrane Compatibility Nitrocellulose, PVDF
Product Line SuperSignal
Quantity 100 mL + 20 mL bundle
Substrate Horseradish Peroxidase
Sensitivity Low-femtogram (mid-zeptomole)
Sufficient For 12 Mini-Gel Blots, 1000 sq. cm of Membrane; 2 Mini-Gel Blots, 200 sq. cm of Membrane
Content And Storage • 100 mL SuperSignal West Femto reagent, store at 2–8°C
• 20 mL SuperSignal West Atto reagent, store at 2–8°C
Shipping Condition Approved for shipment at Room Temperature or on Wet or Dry Ice
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Frequently Asked Questions (FAQs)

How do I handle saturated or burned out bands using SuperSignal West Atto Ultimate Sensitivity Substrate?

Here are some suggestions:
- Shorten the time of exposure
- Wash blot and try using SuperSignal West Pico PLUS Chemiluminescent Substrate
- Strip the membrane using Restore Western Blot Stripping Buffer, re-probe with lesser amount of primary and secondary antibodies, and try first with SuperSignal West Pico PLUS Chemiluminescent Substrate
- Re-run the gel using lesser sample
How can I reduce background with SuperSignal West Atto Ultimate Sensitivity Substrate?

Here are potential causes for high background and corresponding solutions:
- Too much HRP in the system: Further dilute the HRP-conjugate, see guidelines in Table 1 in the user guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0018858_SuperSignalWestAtto_UG.pdf).
- Inadequate blocking: Optimize blocking conditions.
- Inappropriate blocking reagent: Try a different blocking reagent. Use clear milk or StartingBlock blocking buffer instead of dry milk.
- Inadequate washing: Increase length, number or volume of washes.
- Film has been overexposed: Decrease exposure time or use Pierce Background Eliminator (Cat No. 21065).
- Concentration of antigen or antibody is too high: Decrease amount of antigen or antibody.
- Poor antibody specificity: Use SuperSignal Western Blot Enhancer (Cat. No. 46640).
We also recommend minimizing handling of the membrane, using clean gloves while handling, clean plasticware, glassware, forceps, or other contact surfaces. We also recommend trimming of the membrane to remove excess area.
Is SuperSignal West Atto Ultimate Sensitivity Substrate compatible with Restore Western Blot Stripping Buffer?

SuperSignal West Atto Ultimate Sensitivity Substrate is compatible with Restore Western Blot Stripping Buffer.
When should I use SuperSignal West Atto Ultimate Sensitivity Substrate?

We recommend using SuperSignal West Atto Ultimate Sensitivity Substrate for detection of proteins in the following situations:
- For proteins with low concentration, not measurable by BCA or Bradford assays
- For low-abundance phosphorylated or glycosylated proteins
- For low-abundance membrane or nuclear proteins
- For low-abundance proteins being detected with a low-affinity antibody or when the antibody is precious
- When sample source is limited or when multiple conditions or assays are being tested on a single sample

Is SuperSignal West Atto Ultimate Sensitivity Substrate affected by freeze/thaw cycles?

SuperSignal West Atto Ultimate Sensitivity Substrate can tolerate up to 3 freeze/thaw cycles without significant loss in performance.
What is the detection range for SuperSignal West Atto Ultimate Sensitivity Substrate?

SuperSignal West Atto Ultimate Sensitivity Substrate is an ultra-sensitive enhanced chemiluminescent (ECL) substrate that can detect low-femtogram to high-attogram quantities of protein.

For Research Use Only. Not for use in diagnostic procedures.