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Image-iT™ FX Signal Enhancer
Image-iT™ FX Signal Enhancer
Invitrogen™

Image-iT™ FX Signal Enhancer

Image-iT™ FX signal enhancer is a liquid that is applied directly to slides or coverslips containing fixed and permeabilized cellRead more
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Catalog NumberQuantity
I36933Promo Image10 mL
Catalog number I36933
Price (USD)
178.65
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10 mL
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Price (USD)
178.65
Online Exclusive
194.00
Save 15.35 (8%)
Each
Add to cart
Ask our AI about this Product
Image-iT™ FX signal enhancer is a liquid that is applied directly to slides or coverslips containing fixed and permeabilized cell or tissue samples prior to staining with fluorescent probes. When Image-iT™ FX signal enhancer is used, nonspecific fluorescence (background) commonly seen with the application of fluorescent conjugates of streptavidin, goat anti-mouse, or goat anti-rabbit IgG is largely eliminated.

Key Attributes

Easy protocol—apply enhancer from ready-made solution in dropper bottle
Effective with many fluorescent labels (see Table 2 in user manual below)
Cost effective—supplies enough for 50 standard coverslips
Compatible with your immunostaining protocol’s other blocking steps

Reduce Nonspecific Binding of Dyes to Tissues.
Alexa Fluor™ dyes and many other dyes have negatively charged modifications that can lead to nonspecific association with cells and tissue. While not always significant, it can be a problem when pushing the limits of sensitivity in low-abundance targets. Image-iT™ FX signal enhancer can significantly reduce this binding, resulting in marked improvements in staining.

Need help deciding what product you need for your cell imaging? See our cell imaging workflow and decision tree.

Tired of poor signal-to-noise? See our collection antifade and signal enhancer products from Molecular Probes™.

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Product LineImage-iT™
Quantity10 mL
Shelf LifeAt least 6 Months
Shipping ConditionRoom Temperature
Product TypeSignal Enhancer
Solution TypeSignal Enhancer
Unit SizeEach
Contents & Storage
Store at room temperature.
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Frequently asked questions (FAQs)

I used Image-iT FX Signal Enhancer solution to get rid of nonspecific nuclear labeling with Alexa Fluor 568 secondary antibody, but I also saw a significant reduction in my specific mitochondrial antibody labeling. Why is this and what can I do?

The Image-iT FX Signal Enhancer reduces non-specific binding of dye conjugates by blocking positively-charged areas of cells or tissues that attract negatively-charged dyes. In cells after fixation, some positively-charged structures are the nuclei and mitochondria. Thus, you would expect to see a reduction in both mitochondrial and nuclear signal. The lower signal you see afterward is the specific mitochondrial signal; the fluorescence that was lost was the non-specific labeling.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I use the Image-iT FX Signal Enhancer instead of my normal blocking solution (BSA or serum)?

No. Image-iT FX Signal Enhancer is not a protein blocker, like BSA, normal serum, or other commercial antibody blockers. Use it as a separate step to block non-specific charge-based binding of dyes to cellular components.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I am labeling fixed and permeabilized cultured cells with an Alexa Fluor secondary antibody. My secondary-only control is showing nuclear and mitochondrial labeling, even though I did a thorough protein blocking and tried a concentration range. What can be done to minimize this non-specific binding?

This is most likely charge-based binding due to interactions of the charge on the dyes with cellular components of opposite charge. This can be blocked by using the Image-iT FX Signal Enhancer Solution which eliminates non-specific binding due to charge. The Signal Enhancer is applied as a pre-blocking step and your regular blocking regimen should be used to limit non-specific binding due to protein-protein interactions. Signal Enhancer cannot be used on live cells.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I am observing high background when I analyze my click-labeled samples. What is causing this and what can I do to reduce the background?

The click reaction is very selective between an azide and alkyne. No other side reactions are possible in a biological system. Any non-specific background is due to non-covalent binding of the dye to various cellular components. The Select FX Signal Enhancer is not effective at reducing non-specific charge-based binding of dyes following the click reaction; we do not recommend its use with the Click-iT detection reagents. The best method to reduce background is to increase the number of BSA washes. You should always do a no-dye or no-click reaction control under the same processing and detection conditions to verify that the background is actually due to the dye and not autofluorescence. You can also perform the complete click reaction on a carrier solvent-only, no EdU or no-EU control to verify the specificity of the click reaction signal.

Find additional tips, troubleshooting help, and resources within our Cell Viability, Proliferation, Cryopreservation, and Apoptosis Support Center.

I used a neuron-specific antibody to label my neurons. How can I reduce non-specific antibody binding?

A blocking step should be performed to reduce fluorescence due to non-specific antibody binding. A common blocking step is the addition of a 2-5% solution of bovine serum albumin (fraction V defatted BSA). Another approach employs the addition of a 5-10% solution of serum from the species in which the secondary antibodies were raised. For example, when using goat anti-mouse IgG secondary antibodies, samples may be effectively blocked with 5-10% normal goat serum. To further reduce background fluorescence, the Image-iT FX Signal Enhancer can be included as a pre-blocking step to decrease non-specific labeling due to charge interactions between the dyes on the conjugates and the cellular constituents.
If you are using a secondary antibody make sure that the species of the antibody is not the same as the species of the sample. For example do not use an anti-mouse secondary antibody on mouse tissue.
Titrate the antibody to the lowest concentration you can use and still get adequate signal.
Try using a fluorescently tagged primary antibody because it should give reduced background but be aware this can reduce signal intensity.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

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Lot #Certificate TypeDateCatalog Number(s)
3256509Certificate of AnalysisJun 29, 2025I36933
2896700Certificate of AnalysisApr 03, 2024I36933
2795285Certificate of AnalysisNov 19, 2023I36933
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Citations & References (28)

Citations & References
Abstract
NKT cells are critical to initiate an inflammatory response after Pseudomonas aeruginosa ocular infection in susceptible mice.
Authors:Hazlett LD, Li Q, Liu J, McClellan S, Du W, Barrett RP
Journal:J Immunol
PubMed ID:17617607
'CD4(+) T cells produce IFN-gamma contributing to corneal perforation in C57BL/6 (B6) mice after Pseudomonas aeruginosa infection. To determine the role of NK and NKT cells, infected corneas of B6 mice were dual immunolabeled. Initially, more NKT than NK cells were detected, but as disease progressed, NK cells increased, while ... More
Identification of STAT3 as a substrate of receptor protein tyrosine phosphatase T.
Authors:Zhang X, Guo A, Yu J, Possemato A, Chen Y, Zheng W, Polakiewicz RD, Kinzler KW, Vogelstein B, Velculescu VE, Wang ZJ,
Journal:Proc Natl Acad Sci U S A
PubMed ID:17360477
'Protein tyrosine phosphatase (PTP) receptor T (PTPRT) is the most frequently mutated PTP in human cancers. However, the cell signaling pathways regulated by PTPRT have not yet been elucidated. Here, we report identification of signal transducer and activator of transcription 3 (STAT3) as a substrate of PTPRT. Phosphorylation of a ... More
Displacement of SERCA from SR lipid caveolae-related domains by Bcl-2: a possible mechanism for SERCA inactivation.
Authors:Dremina ES, Sharov VS, Schöneich C
Journal:Biochemistry
PubMed ID:16388593
'Bcl-2 exerts its anti-apoptotic effect in part through the regulation of Ca2+ homeostasis at the level of the endoplasmic reticulum. Earlier, we demonstrated that a truncated form of Bcl-2, Bcl-2delta21, interacts with and destabilizes the skeletal muscle sarco/endoplasmic reticulum Ca-ATPase (SERCA) [Dremina, E. S., Sharov, V. S., Kumar, K., Zaidi, ... More
Androgen induces expression of the multidrug resistance protein gene MRP4 in prostate cancer cells.
Authors:Cai C, Omwancha J, Hsieh CL, Shemshedini L
Journal:Prostate Cancer Prostatic Dis
PubMed ID:17003774
'Multidrug resistance-associated proteins (MRPs) may mediate multidrug resistance in tumor cells. Using a gene array analysis, we have identified MRP4 as an androgen receptor (AR)-regulated gene. Dihydrotestosterone induced MRP4 expression in both androgen-dependent and -independent LNCaP cells, whereas there was little detectable expression in PC-3 or normal prostate epithelial cells. ... More
Constitutive interferon-inducible protein 16-inflammasome activation during Epstein-Barr virus latency I, II, and III in B and epithelial cells.
Authors:Ansari MA, Singh VV, Dutta S, Veettil MV, Dutta D, Chikoti L, Lu J, Everly D, Chandran B,
Journal:
PubMed ID:23720728
'Epstein-Barr virus (EBV), etiologically linked with human B-cell malignancies and nasopharyngeal carcinoma (NPC), establishes three types of latency that facilitate its episomal genome persistence and evasion of host immune responses. The innate inflammasome responses recognize the pathogen-associated molecular patterns which lead into the association of a cytoplasmic sensor such as ... More
28 total citations

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