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  5. Thermo Scientific™ SuperSignal™ ELISA Femto Substrate

Thermo Scientific™ SuperSignal™ ELISA Femto Substrate

Product Code. 11879310 Shop All Thermo Scientific Products
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Product Code. 11879310 Supplier Thermo Scientific™ Supplier No. 37075

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Description

HRP substrate formulated for superior protein detection and low-end linearity in chemiluminescent ELISA applications.

Thermo Scientific™ SuperSignal ELISA Femto Maximum Sensitivity Substrate is formulated for superior protein detection and low-end linearity in chemiluminescent ELISA applications.

SuperSignal ELISA Femto Substrate generates detectable chemiluminescence (light) within one minute of addition to trace amounts of soluble HRP, saving up to 30 minutes per assay compared to colorimetric ELISA substrates. This feature is ideal for high-throughput screening (HTS) applications in which as many as 100,000 assays can be performed daily on robotic equipment. Because incubation periods are often a limiting factor in the development of rapid automated assays, SuperSignal ELISA Femto Substrate is the logical choice for automated HTS applications.

Highlights:

  • Immediate light generation – intense signal generated immediately at both room temperature and 37°C

  • Improved low-end linearity – easy detection of low quantities of proteins with high signal-to-noise ratios and low-end linearity of dose response curves

  • High sensitivity – femtogram-level detection of target proteins

  • Reduction in assay time – high sensitivity allows for reduction in incubation steps

  • Stability – storage for six months at room temperature or a minimum of 12 months at 4°C with a six-hour working solution stability

  • Emits light at 425nm – measurable by luminometers

Includes:

Luminol/Enhancer Solution; Peroxide Buffer

TRUSTED_SUSTAINABILITY

Specifications

Product Type ELISA Substrate
Quantity 100 mL Kit
Detection Method Chemiluminescence
Product Line SuperSignal
Substrate Properties Chemical Substrate
Substrate Type HRP (Horseradish Peroxidase)
Content And Storage Stable for one year at 4°C or six months at room temperature. Product is shipped at ambient temperature.

Frequently Asked Questions (FAQs)

What type of instrument do I use to read the plates detected with the SuperSignal ELISA Femto Maximum Sensitivity Substrate?

A luminometer is used to measure relative light units.
What microplates work best with the SuperSignal ELISA Femto Maximum Sensitivity Substrate?

White plates provide the best signal-to-noise ratio. Black plates provide lower background but less detectable signal. Here are two suggestions we offer for white plates:

-Pierce 96-Well Polystyrene Plates, White Opaque (Cat. No. 15042)
-Nunc FluoroNunc/LumiNunc 96-Well Plates (Cat. No. 437796)

What signal duration occurs with SuperSignal ELISA Femto Maximum Sensitivity Substrate?

Maximum signal is reached within two minutes and then declines steadily until it is neglible after twenty minutes.
How sensitive is SuperSignal ELISA Femto Maximum Sensitivity Substrate?

Our researchers have detected down to 168 fg of interleukin-2 in sandwich ELISA.
Can I dilute the SuperSignal ELISA Femto Maximum Sensitivity Substrate working reagent if my signal is too intense?

We do not recommend diluting the working reagent. It is better to dilute or otherwise optimize primary and or secondary antibody concentrations or use more washes between incubation steps.
At what wavelength is SuperSignal ELISA Femto Maximum Sensitivity Substrate measured?

The light emission wavelength is measured at 425 nm.
Can I use SuperSignal West Dura Extended Duration Substrate with imaging systems?

Yes. The SuperSignal Western blot substrates are well suited to most imaging systems including CCD camera units; however, most screen-type phosphorimagers give poor results with the SuperSignal substrates.
Can I "fix" my overexposed film when using SuperSignal West Dura Extended Duration Substrate?

We offer the Pierce Background Eliminator Kit (Cat. No. 21065) that reduces high background, shading, speckling, and over-exposed bands on exposed film.
How can I increase detection sensitivity with SuperSignal West Dura Extended Duration Substrate?

Treating the nitrocellulose or PVDF membrane with Western Blot Signal Enhancer (Cat. No. 21050) before blocking can boost the chemiluminescent signal of low-intensity bands.
Can the quality of the membrane affect the detection results when using SuperSignal West Dura Extended Duration Substrate?

A high-quality membrane can effectively bind more target protein and result in a lower background after detection when using SuperSignal substrates.
Can I strip and reprobe my western blot when using SuperSignal West Dura Extended Duration Substrate?

Nitrocellulose and PVDF membranes can be stripped following detection with SuperSignal chemiluminescent substrates, for reprobing. Restore Western Blot Stripping Buffer (Cat. No. 21059, 21063) can be used to gently but efficiently remove probes from the western blot membrane without damaging the target protein. In most cases, re-blocking of the membrane is not necessary.
I am using SuperSignal West Dura Extended Duration Substrate. Why is my signal disappearing after only a few minutes?

The most common cause of premature signal burnout is the use of excessive peroxidase-conjugated probe. During the substrate reaction, an extreme excess of peroxidase will deplete the limited amount of substrate resulting in intense, but short, signal duration. To avoid signal fading, it is imperative to use the peroxidase conjugated probe at a concentration recommended in the SuperSignal West Dura Extended Duration Substrate manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0011307_SupSig_West_Dura_Extend_Dur_Subs_UG.pdf).
How can I increase the signal intensity when I am using SuperSignal West Dura Extended Duration Substrate?

The following are possible causes for weak signal intensity:
- Low level of protein transferred from the gel to the membrane. This is particularly true for high molecular weight proteins. Reversible Protein Stain Kits for Nitrocellulose and PVDF membranes (Cat. Nos. 24580 and 24585, respectively) may be used to stain the bands on the membrane to indicate transfer efficiency.
- Over-diluted primary and secondary antibody probes. We suggest using the recommended dilution for the antibody probes for each substrate. More sensitive substrates will require a more dilute antibody solution.
- Short substrate incubation time. For best results, incubate the membrane for 5-15 min in the SuperSignal substrate.
Have you tested the SuperSignal ELISA Femto Substrate with magnetic beads?

We have not tested the SuperSignal ELISA Femto Substrate with magnetic beads. However, we know that some metals can interfere with the substrate. Therefore, depending on the chemistry of the beads, they may or may not interfere with the substrate.
Is the time taken to add your SuperSignal ELISA Femto Substrate to the wells of a microplate important?

Yes. The signal from the SuperSignal ELISA Femto Substrate ramps up very quickly and optimal signal production lasts only for 5-10 min. Therefore, we recommend using a multichannel pipette to add the substrate to the wells, mix for 1 min, and then read the signal produced. Typically, reading each well for 0.1-1 sec will provide good signal with this substrate.
Note: With the SuperSignal ELISA Pico Chemiluminescent Substrate (Cat. No. 37070, 37069), the signal does not decay as quickly and researchers typically have up to 30 min to read the plate.

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

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