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Pierce Immunostain Enhancer


Alleviates immunostaining problems such as low signal and poor sensitivity.

Pierce Immunostain Enhancer

Thermo Scientific Pierce Immunostain Enhancer enables significant antibody dilution (5- to 20-fold beyond normal) to increase signal intensity 3- to 12-fold in immunostaining applications with chromogenic or fluorescent substrates.

This ready-to-use enhancer for immunohistochemical staining does not add steps to the workflow because it is used to dilute the primary and secondary antibodies. The Pierce Immunostain Enhancer is compatible with fluorescence and chromogenic detection and routinely increases both signal intensity and detection sensitivity. Signal enhancement is antibody-dependent and typically ranges from 3- to 12-fold. Because of the strong signal enhancement, the Pierce Immunostain Enhancer reduces the amount of antibody required to achieve optimal detection.

Highlights:

  • Save precious antibody – Pierce Immunostain Enhancer allows the customer to use only a fraction of antibody to achieve the same signal as with conventional immunodetection
  • Convenience – simply replace your current antibody dilution buffer with Pierce Immunostain Enhancer (unlike other signal enhancement methods which require additional steps)
  • Increased signal intensity and sensitivity – provides 3- to 12- fold increase in signal intensity and sensitivity for improved visualization of the antigen of interest in cells and tissues
  • Improved specificity – significantly improves signal-to-noise ratio for poor quality and low affinity antibodies
  • Compatible – can be used with chromogenic and fluorescent detection methods

Product Details:

The Pierce Immunostain Enhancer enables significant dilution of primary and secondary antibodies. Dilution 5- to 20-fold beyond the antibody vendor’s recommendation has been used successfully; however, the dilution factor varies depending on the antibody, tissue, and antigen.

Pierce Immunostain Enhancer enables a 200-fold dilution of Lamin B1 primary antibody beyond the vendor recommendation.
Save money by using only a fraction of your precious primary antibody by diluting in Thermo Scientific Pierce Immunostain Enhancer (200-fold dilution of Lamin B1 primary antibody beyond the vendor’s recommendation). A549 cells were seeded at 5000 cells per well in a 96-well plate and incubated for 18-20 hours at 37°C, 5% CO2 in a humidified incubator. The cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Thermo Scientific Surfact-Amps X-100 and then blocked at room temperature (RT) for 30 minutes with 2% BSA Blocker containing 0.1% Triton™ X-100 (standard buffer). The primary and the secondary antibodies were diluted with either the standard buffer or with Thermo Scientific Pierce Immunostain Enhancer. A549 cells were probed with rabbit polyclonal anti-lamin B1 for 1 hour at room temperature, and then the lamin B1 was detected with Thermo Scientific DyLight 594-conjugated goat anti-rabbit antibody (4µg/mL) (Part No. 35560). Lamin B1 antibody specifically stains the nuclear envelope. Panel A: Lamin B1 is detected in A549 cells using a 1:125 dilution of the primary antibody in BSA; Panel B: The same dilution results in oversaturation of the signal when the Pierce Immunostain Enhancer is used. Panel C: The antigen is undetectable when the primary antibody is diluted 1:25,000 in standard buffer; Panel D: Pierce Immunostain Enhancer results in clear detection of lamin B1 at the same dilution as Panel C. Images were acquired at the same gain and exposure time using appropriate optical filter sets with a 20X (0.45 NA) objective.

Pierce Immunostain Enhancer enables a 25 times less less Ezrin primary antibody to be used for improved signal.
Increase sensitivity and signal intensity for fluorescent detection of specific targets when using Thermo Scientific Pierce Immunostain Enhancer. A549 cells were seeded at 5000 cells per well in a 96-well plate and incubated for 18-20 hours at 37°C, 5% CO2 in a humidified incubator. A549 cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Thermo Scientific Surfact-Amps X-100 and then blocked at room temperature (RT) for 30 minutes with 2% of Thermo Scientific Blocker BSA (Part No. 37525) + 0.1% Triton™ X-100). The primary and the secondary antibodies were diluted with either a standard buffer or the Pierce Immunostain Enhancer. A549 cells were probed with mouse monoclonal anti-Ezrin; incubated for 1 hour at room temperature and then detected with Thermo Scientific DyLight 488 conjugated to goat anti-mouse antibody (4µg/mL) (Part No. 35502). Ezrin antibody specifically stains microvillar peripheral membrane protein in the cytoplasm (see arrows). Left panel: Detection of ezrin in standard buffer (dilution 1:200). Right panel: Using 25-times less Ezrin antibody (dilution 1:1250) in Pierce Immunostain Enhancer shows improved sensitivity and better visualization of the antigen of interest in the cells. Images were acquired at the same gain and exposure time using appropriate optical filter sets with a 20X (0.45 NA) objective.

Pierce Immunostain Enhancer is compatible with chromogenic detection methods (Metal Enhnaced DAB) for detection of Cytokaratin 18 in adenocarcinoma tissue.
Improve Specificity, Signal Intensity and Reduce background using Thermo Scientific Pierce Immunostain Enhancer in chromogenic detection Immunohistochemistry. Formalin-fixed paraffin embedded poorly differentiated colon adenocarcinoma tissue section was deparaffinized then subjected to heat-induced epitope retrieval using citrate buffer, followed by endogenous peroxidase quenching and then blocking with 2% BSA Blocker containing 0.1% Triton™ X-100 (standard buffer). The primary and the secondary antibodies were both diluted with either the standard buffer or the Pierce Immunostain Enhancer. The tissue was incubated with rabbit anti-cytokeratin 18 at 0.01µg/mL for 1 hour at room temperature. After washing with ultrapure water containing 0.05% Tween™ 20 Detergent, the tissues were incubated with HRP-conjugated goat anti-rabbit antibody at 1µg/mL for 45 minutes (room temperature). Cytokeratin was revealed by chromogenic detection using Metal Enhanced DAB with peroxide substrate (Part No. 34065). The images were acquired with a 20X/0.45NA objective on Zeiss Axio Observer™ .Z1 using an Axio MRC3 color Digital Camera. Cytokeratin 18 detection is dramatically increased using the enhancer. Additional benefits were found in the reduction of extraneous background information, as well as, stronger signal intensity and sensitivity due to the much shorter development of the chromogenic substrate.

Pierce Immunostain Enhancer is compatible with fluorescent detection of CEA in formalin-fixed, paraffin embedded poorly differentiated colon adenocarcinoma tissue section.
Improve specificity and signal intensity using Thermo Scientific Pierce Immunostain Enhancer for fluorescent immunohistochemistry. Formalin-fixed paraffin embedded poorly differentiated colon adenocarcinoma tissue section was deparaffinized then subjected to heat-induced epitope retrieval using citrate buffer, followed by blocking with 2% BSA Blocker + 0.1% Triton™ X-100 (standard buffer). The primary and the secondary antibodies were diluted with either the standard buffer or with Pierce Immunostain Enhancer. The tissue was incubated with rabbit anti-CEA antibody at 0.1µg/mL for 1 hour at room temperature then detected with DyLight 488-conjugated goat anti-rabbit antibody at 4µg/mL for 45 minutes (room temperature). The images were acquired at the same gain and exposure time with a 20X/0.45NA objective on Zeiss Axio Observer™ Z1 Fluorescence Microscope using an ORCA-ER-1394 Hamamatsu, CCD Digital Camera.

Related Resources:

Immunohistochemistry (IHC) technical guide
IHC troubleshooting technical guide

Related Products:

All Immunohistochemistry Reagents


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
46645 Pierce Immunostain Enhancer
Formulation: Proprietary solution
Sufficient For: 10 large (3 square cm each) tissue section slides
2mL Product Instructions for product #46645 Pierce Immunostain Enhancer MSDS for product #46645 Pierce Immunostain Enhancer Certificate of Analysis for product #46645 Pierce Immunostain Enhancer $45.00

Add to cart
46644 Pierce Immunostain Enhancer
Formulation: Proprietary solution
Sufficient For: 100 large (3 square cm each) tissue section slides
20mL Product Instructions for product #46644 Pierce Immunostain Enhancer MSDS for product #46644 Pierce Immunostain Enhancer Certificate of Analysis for product #46644 Pierce Immunostain Enhancer $318.00

Add to cart
46644B Pierce Immunostain Enhancer
Formulation: Proprietary solution
Custom MSDS for product #46644B Pierce Immunostain Enhancer Certificate of Analysis for product #46644B Pierce Immunostain Enhancer

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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