Thermo Scientific™ SuperSignal™ West Atto Ultimate Sensitivity Substrate

Catalog No.	Quantity
PIA38555	100 mL
PIA38554	20 mL
PIA38556	200 mL

Catalog No. PIA38555 Supplier Thermo Scientific™ Supplier No. A38555

Description

SuperSignal™ West Atto Ultimate Sensitivity Substrate is an enhanced chemiluminescent (ECL) substrate for high attogram protein detection and provides the high level sensitivity and better signal-to-noise ratios compared to other commercially available horseradish peroxidase (HRP) substrates.

• Sensitivity: low-femtogram to high-attogram
• Stability: eight hours for working solution, one year for kit at 4°C
• Compatibility: nitrocellulose and PVDF membranes
• Signal duration: up to four hours
• Recommended primary antibody concentration: 1:1000 to 1:5000 dilution (0.2 to 1.0 μg/mL)
• Recommended secondary antibody concentration: 1:100000 to 1:250000 dilution (4 to 10 ng/mL)

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
• Substrate: Horseradish Peroxidase
• Sensitivity: Low-femtogram to high-attogram
• Sufficient For: 12 Mini-Gel Blots, 1000 sq. cm of Membrane
• Kit Contents: Luminol/Enhancer, 50 mL, Stable Peroxide Buffer, 50 mL
• Content And Storage: SuperSignal West Atto Substrate / SuperSignal West Atto Stable Peroxide
  
  • Substrate Solution, 50 mL
  • Stable Peroxide Solution, 50 mL
  
  Sufficient for 12 mini-gel blots; 1000 cm² of membrane.
  
  Store reagents at 2–8°C.
• Shipping Condition: Wet Ice

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.