TSA® Research Reagents

MANUALResearch Use Only. Not for use in diagnostic procedures.

TSA Fluorescence Systems

Material Provided
Format*TSA
Tetramethylrhodamine

TSA Coumarin

TSA Cyanine 3

Kit Components

NEL701A001KT

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1X Amplification Diluent (15 mL)
Fluorescein Tyramide (dry, dissolve in 300 µL DMSO)Streptavidin-HRP (150 µL)
Blocking Reagent (3 g)

100-300 slides

NEL701001KT

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1X Amplification Diluent (30 mL)
Fluorescein Tyramide (dry, dissolve in 600 µL DMSO)Streptavidin-HRP (2 x 150 µL)
Blocking Reagent (10 g)

100-300 slides

SAT701001EA••

1X Amplification Diluent (30 mL)
Fluorescein Tyramide (dry, dissolve in 600 µL DMSO)

500-1500 slides SAT701B001EA••

1X Amplification Diluent (2 x 75 mL)
Fluorescein Tyramide (5 tubes, dry, dissolve in 600 µL DMSO/tube)

100-300 slides

NEL702001KT

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1X Amplification Diluent (30 mL)
Tetramethylrhodamine Tyramide (dry, dissolve in 600 µL DMSO)Streptavidin-HRP (2 x 150 µL)
Blocking Reagent (10 g)

100-300 slides

SAT702001EA

100-300 slides

NEL703001KT

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1X Amplification Diluent (30 mL)
Tetramethylrhodamine Tyramide (dry, dissolve in 600 µL DMSO)
1X Amplification Diluent (30 mL)
Coumarin Tyramide (dry, dissolve in 600 µL DMSO)Streptavidin-HRP (2 x 150 µL)
Blocking Reagent (10 g)

50-150 slides

NEL704A001KT

50-150 slides

SAT704A001EA

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1X Amplification Diluent (15 mL)
Cyanine 3 Tyramide (dry, dissolve in 300 µL water)
Streptavidin-HRP (150 µL)
Blocking Reagent (3 g)
1X Amplification Diluent (15 mL)
Cyanine 3 Tyramide (dry, dissolve in 300 µL water)
1X Amplification Diluent (75 mL)
Cyanine 3 Tyramide (5 tubes, dry, dissolve in 300 µL water)
1X Amplification Diluent (15 mL)
Cyanine 5 Tyramide (dry, dissolve in 300 µL water)
Streptavidin-HRP (150 µL)
Blocking Reagent (3 g)••

1X Amplification Diluent (15 mL)
Cyanine 5 Tyramide (dry, dissolve in 300 µL water)

50-150 slides

TSA Fluorescein

Catalog #250-750 slides SAT704B001EA
50-150 slides

NEL705A001KT

50-150 slides

SAT705A001EATSA Cyanine 5*The format of the kit is based on 100 - 300 µL per slide of Fluorophore Tyramide Working Solution (see page 3).

Product Information
Storage:
Stability:

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Store kits in the dark at 4ºC.
The components in the kits are stable for a minimum of 3 months under proper storage
conditions.

Application:

TSA Fluorescence kits are intended for high sensitivity detection in immunohistochemistry(IHC), immunocytochemistry (ICC) and in situ hybridization (ISH) experiments. Final detection
may be fluorescent or chromogenic (with TSA Fluorescein kits only).
FOR RESEARCH USE ONLY.

Safety Note

All reagents are classified as nonhazardous. However, it is strongly recommended to wear
disposable gloves and safety glasses while working. Thorough washing of hands after handling is
also recommended.

Quality Control

We certify that QC results of these reagents meet our quality release criteria.

What is TSA technology?
Tyramide Signal Amplification (TSA™) from PerkinElmer is a technology that improves sensitivity by up to 100-foldsensitivity while allowing reduced consumption of primary antibodies or probes.
TSA Fluorescence Kits use horseradish peroxidase (HRP) to catalyze covalent deposition of fluorophores directly
adjacent to the immobilized enzyme. The labeling reaction is quick (less than 10 minutes) and deposited labels can thenbe viewed directly using standard or confocal microscopy. TSA Fluorescein may also be used in combination with antifluorescein enzyme conjugates and appropriate chromogenic substrates for brightfield microscopy.
The use of TSA reagents results in a significant increase in sensitivity over standard detection methods, while maintaining
specificity and resolution. Moreover, TSA reagents allow drastically reduced consumption of primary antibody or probe.

Reagents and Materials
Critical Reagents Required but not Supplied• Depending upon your assay, additional HRP-labelled reagent to drive the signal amplification reaction may berequired. For example:o Anti-digoxigenin-HRP for use with digoxigenin labeled probes or antibodies (cat. no. NEF832001EA)o Anti-fluorescein-HRP for use with fluorescein labeled probes or antibodies (cat. no. NEF710001EA)o HRP-conjugated anti-species secondary antibody
 Anti-rabbit IgG (goat) HRP (cat. no. NEF812001EA)
 Anti-mouse IgG (goat) HRP (cat. no. NEF822001EA)• DMSO (molecular biology or HPLC grade)• Buffer components and detergents such as PBS, Triton-X100, Tween-20• Blocking Reagent (cat. no. FP1012 or equivalent)

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Solutions to prepare
The following buffers and reagents are required for slide preparation and signal amplification
Fluorophore Tyramide Stock Solution
Fluorophore Tyramide Reagent is supplied as a solid. Each vial must be reconstituted with DMSO or water as indicated tomake Fluorophore Tyramide Stock Solution. Fluorophore Tyramide Stock Solution is stable for at least 3 months when stored
at 4ºC. (Note: DMSO freezes at 4ºC; therefore, thaw Stock Solution before each use). Here are recommended dissolution
volumes for stock solutions.
Catalog #Tyramide Stock Solution Preparation
NEL701A001KT
Add 300 µL DMSO to reagent tube
TSA Fluorescein

NEL701001KT

SAT701001EA
SAT701B001EA
NEL702001KT
TSA Tetramethylrhodamine
SAT702001EA
NEL703001KTTSA Coumarin
NEL704A001KTTSA Cyanine 3
SAT704A001EA
SAT704B001EA
NEL705A001KTTSA Cyanine 5
SAT705A001EA

Add 600 µL DMSO to reagent tube
Add 600 µL DMSO to reagent tube
Add 600 µL DMSO to reagent tube
Add 600 µL DMSO to reagent tube
Add 600 µL DMSO to reagent tube
Add 600 µL DMSO to reagent tube
Add 300 µL water to reagent tube
Add 300 µL water to reagent tube
Add 300 µL water to reagent tube
Add 300 µL water to reagent tube
Add 300 µL water to reagent tube

TNT Wash Buffer
0.1 M TRIS-HCl, pH 7.5
0.15 M NaCl
0.05% Tween®20
Other wash buffers (such as PBS) may be used. Substitution of 0.3% Triton X-100 for the 0.05% Tween-20 is also possible.
Users should validate the use of alternative wash buffers with their own systems.
TNB Blocking Buffer
0.1 M TRIS-HCl, pH 7.5
0.15 M NaCl
0.5% Blocking Reagent (available separately, catalog number FP1012)
Add Blocking Reagent slowly in small increments to buffer while stirring. Heat gradually to 55°C with continuous stirring tocompletely dissolve the Blocking Reagent. (This should take no longer than 30-60 minutes.) The solution will appear milky.
Bring to room temperature before using. Aliquot and store at -20°C for long term use.
Fluorophore Tyramide Working Solution
Before each procedure, dilute Fluorophore Tyramide Stock Solution1:50 in 1X Amplification Diluent to make Fluorophore
Tyramide Working Solution. Approximately 100-300 µL of Fluorophore Tyramide Working Solution is required per slide.
Discard any unused portion of Fluorophore Tyramide Working Solution.

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Recommendations
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Do not let slides dry out between steps.
A humidified chamber is recommended for all incubation steps (i.e., a damp paper towel in a covered box).
Drain off as much of the incubation solutions as possible, before the addition of the next solution, to prevent reagent
dilution and uneven staining. Blot area around, but not on, tissue section using a tissue.
Be sure to use enough volume of each reagent to completely cover sections or cells.
Optional: Use of coverslips may reduce reagent evaporation, especially during steps which require long incubation atelevated temperatures (such as probe hybridization). However care must be taken upon removal to prevent damage
to tissues or cells.
If your assay includes streptavidin conjugates, check for endogenous biotin which may be a source of non-specificbackground.
If there is too much signal, dilute the primary antibody, probe or HRP conjugate further. TSA kits are designed foruse at 1:50 dilution of the tyramide reagent for optimal results.
First time users should apply TSA to a proven system.

Quenching Endogenous Peroxidase
Activation and covalent binding of TSA reagent is catalyzed by peroxidase. Endogenous peroxidase will be a source of
background, and should be quenched if present. Users should establish the need for doing this and optimal methodology
specific to the tissues or cells being stained. Options include:• 0.3% H2O2 to 3% H2O2 in PBS, incubation for 10 to 60 minutes• 0.3% H2O2 to 3% H2O2 in Methanol, incubation for 10 to 60 minutes
For paraffin-embedded tissues, quenching can be done after dewaxing and alcohol rehydration but before the protease
digestion step. For frozen tissue or cell preps, quenching can be done following fixation and before the protease
digestion step. After quenching wash with TNT or 1X PBS buffer for 5 minutes. See Li et al from references section for
more suggestions.

TSA-ISH Optimization
Inadequate optimization of probe and HRP conjugate dilution may be a source of high background and reduced signal.
TSA is ideal for detection of targets that have weak signals using standard detection methods. If the signal is strong withstandard detection methods, dilute the probe until the signal begins to disappear and then proceed with TSA
optimization.
Probe Optimization• Slide 1: use same probe dilution as standard method.
• Slide 2: 5-fold dilution from slide 1• Slide 3: 2-fold dilution from slide 2 (further dilution may be necessary)• Slide 4: negative control (probe omitted)
HRP Conjugate Dilution• SA-HRP included with TSA Systems: 1:100 dilution• Streptavidin-HRP (catalog number NEL750001EA): 1:250-1:1000 dilution• Anti-fluorescein-HRP (catalog number NEF710001EA): 1:100-1:500 dilution• Anti-DNP-HRP (catalog number FP1129): 1:100-1:500 dilution• Anti-digoxigenin-HRP (catalog number NEF832001EA): 1:500-1:5000 dilution• HRP reagents from other manufacturers: Begin with recommended range for slide applications. Further dilution
may be needed

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TSA-ISH Protocol Overview
Standard
nonradiometric
ISH
Blocking Step
Incorporationof HRP
TSA
Amplification

Visualization

Quench endogenous peroxidase activity (if needed)
Tissue permeabilization (if needed)
Probe hybridization
Post-hybridization stringency washes↓
Block slides for 30 min. in TNB buffer at room temperature↓
Incubate slides with appropriate HRP reagent (anti-DIG-HRP, SA-HRP, etc.) for 30 min. at room
temperature↓
Wash slides 3X for 5 min. in TNT buffer at room temperature Incubate in tyramide working solution
for 3 to 10 min. at room temperature↓
Wash slides 3X for 5 min. in TNT buffer at room temperature↓
Fluorescent
Chromogenic (TSA Fluorescein Only)
Counterstain and mount for fluorescence
Microscopy

Incubate slides in anti-fluorescein-HRP or
anti-fluorescein-AP
for 30 min. at room temperature↓
Wash slides 3X for 5 min. in TNT buffer atroom temperature↓
Add appropriate chromogen, counterstain andmount for microscopy

Step by Step TSA-ISH Protocol
Step 1: Slide Preparation
Prepare tissues or cells using standard fixation and embedding techniques. Dewax and rehydrate slides according tostandard procedures.
Step 2: Standard Non-radioactive ISH
Follow standard non-radioactive in situ hybridization techniques. Include tissue permeabilization (if needed) andquenching of endogenous peroxidase activity (if needed). Probe hybridization (with digoxigenin, biotin, DNP, or
fluorescein-labeled probes) should be done using at concentration determined in optimization studies followed by posthybridization stringency washes.
NOTE: Always run an unamplified control slide and an amplified negative control slide with each experiment.
Step 3: Blocking Step
Cover tissue sections with TNB buffer (or other validated blocking buffer) and incubate slides in a humidified chamber for
30-60 minutes at room temperature or at 4ºC overnight.

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Step 4: Introduction of HRP
Incubate slides for 30 minutes at room temperature or at 4ºC overnight in a humidified chamber with appropriate HRPlabeled reagent. Use adequate reagent volume to cover the tissue section, generally 100-300 µL per slide.
• For DIG-labeled probes, use anti-digoxigenin-HRP (catalog number NEF832001EA) diluted in TNB Buffer (orother validated blocking buffer) to concentration determined in optimization studies.
• For biotin-labeled probes, use SA-HRP (catalog number NEL750001EA) diluted in TNB Buffer (or other validated
blocking buffer) to concentration determined in optimization studies.
• For fluorescein-labeled probes, use anti-fluorescein-HRP (catalog number NEF710001EA) diluted in TNB Buffer(or other validated blocking buffer) to concentration determined in optimization studies.
• For DNP-labeled probes, use anti-DNP-HRP (catalog number FP1129) diluted in TNB Buffer (or othervalidated blocking buffer) to concentration determined in optimization studies.
Step 5: Wash
Wash the slides 3X for 5 minutes each in TNT Buffer (or other validated wash buffer) at room temperature with agitation.
Step 6: Amplification
Pipette Fluorophore Tyramide Working Solution onto each slide. Use enough working solution to completely cover thetissue section, generally 100-300 µL per slide. Incubate the slides at room temperature for 3 to 10 minutes.
Step 7: Wash
Wash the slides 3X for 5 minutes each in TNT buffer (or other validated wash buffer) at room temperature with agitation.
Step 8: Visualization of Deposited Fluorophores
For fluorescent detection, counterstain and mount for fluorescence microscopy. (See page 10 table, Fluorophore
Excitation and Emission Maxima.)
For chromogenic detection with TSA Fluorescein, incubate slides with anti-fluorescein-HRP (NEF710001EA) or antifluorescein-AP (NEF709001PK) followed by detection with an appropriate chromogenic substrate.

TSA-IHC Optimization
Inadequate optimization of primary antibody and HRP conjugate dilution may be a source of high background andreduced signal.
TSA is ideal for detection of targets that have weak signals with standard detection methods. It enables use of much
more dilute antibody solutions for improved specificity. If the signal is strong with standard detection methods, dilute theprimary antibody until the signal begins to disappear and then proceed with TSA optimization.
Primary Antibody Optimization• Slide 1: use same antibody dilution as standard method.
• Slide 2: 5-fold dilution from slide 1• Slide 3: 5-fold dilution from slide 2• Slide 4: 5-fold dilution from slide 3 (further dilution may be necessary)• Slide 5: negative control (primary antibody omitted)
HRP Conjugate Dilution• SA-HRP included with TSA Systems: 1:100 dilution• Anti-mouse-HRP (catalog number NEF822001EA): 1:500 to 1:2000 dilution• Anti-rabbit-HRP (catalog number NEF812001EA): 1:500 to 1:2000 dilution• Streptavidin-HRP (catalog number NEL750001EA): 1:1250-1:2500 dilution• Anti-fluorescein-HRP (catalog number NEF710001EA): 1:100-1:500 dilution• HRP reagents from other manufacturers: Begin with recommended range for slide applications. Further dilution
may be needed

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TSA-IHC Protocol Overview
Quench endogenous peroxidase activity (if needed)↓
Block slides for 30 min. in TNB buffer at room temperatureStandard
IHC

Incorporationof HRP

TSA
Amplification

Visualization↓
Incubate slides in primary antibody for 30-60 minutes at room temperature↓
Wash slides 3X for 5 min. in TNT buffer at room temperature↓
Incubate slides in biotinylated secondary
antibody 30-60 min. at room temperature
Incubate slides in HRP labeled secondary
Wash slides 3X for 5 min. in TNT buffer atorroom temperature
antibody for 30 min. at room temperature
Incubate slides in SA-HRP for 30 min. at room
temperature↓
Wash slides 3X for 5 min. in TNT buffer at room temperature↓
Incubate in Fluorophore Tyramide Working Solution for 3 to 10 min. at room temperature↓
Wash slides 3X for 5 min. in TNT buffer at room temperature↓
Fluorescent
Chromogenic (TSA Fluorescein Only)
Counterstain and mount for fluorescence
Microscopy

Incubate slides in anti-fluorescein-HRP or
anti-fluorescein-AP for 30 min. at room
temperature↓
Wash slides 3X for 5 min. in TNT buffer atroom temperature↓
Add appropriate chromogen, counterstain andmount for microscopy

Step by Step TSA-IHC Protocol
Step 1: Slide Preparation
Prepare tissues or cells for detection with TSA using standard fixation and embedding techniques. Dewax and rehydrateusing standard protocols. Quench endogenous peroxidase activity if necessary.
NOTE: Always run an unamplified control slide and an amplified negative control slide with each experiment.
Step 2: Blocking Step
Cover tissue sections with TNB buffer (or other validated blocking buffer) and incubate slides in a humidified chamber for
30-60 minutes at room temperature or at 4ºC overnight.
Step 3: Primary Antibody Incubation
Drain off the blocking buffer and apply primary antibody, diluted in TNB Buffer (or other validated blocking buffer).
Incubate the primary antibody preparation per the manufacturer's instructions regarding incubation time and temperature

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requirements. Use enough volume to completely cover the tissue section (generally 100-300 µL per slide) at theconcentration determined in optimization studies.
Step 4: Wash
Wash the slides 3X for 5 minutes each in TNT Buffer (or other validated wash buffer) at room temperature with agitation.
Step 5: Introduction of HRP
Incubate slides for 30 minutes at room temperature or at 4ºC overnight in a humidified chamber with appropriate HRPlabeled reagent. Use adequate reagent volume to cover the tissue section, generally 100-300 µL per slide. Options
include.
• HRP labeled secondary antibody diluted in TNB Buffer (or other validated blocking buffer).
• 100-300 µL of biotinylated secondary antibody diluted in TNB Buffer (or other validated blocking buffer). Incubate
30-60 minutes in a humidified chamber. Wash the slides for 3 X 5 minutes TNT buffer at room temperature withagitation. Follow by 100-300 µL of SA-HRP diluted in TNB Buffer. Use SA-HRP at 1:100 dilution, or at 1:2000 if
using PerkinElmer Cat. # NEL750001EA.
• When using alternative suppliers, reagents should be optimized for use with TSA starting with manufacturer's
recommended dilutions. Incubate slides in a humidified chamber for 30-60 minutes at room temperature or at
4ºC overnight.
Step 6: Wash
Wash the slides 3X for 5 minutes each in TNT Buffer (or other validated wash buffer) at room temperature with agitation.
Step 7: Amplification
Pipette 100-300 µL of Fluorophore Tyramide Working Solution onto each slide. Incubate the slides at room temperaturefor 3 to 10 minutes.
Step 8: Wash
Wash the slides 3X for 5 minutes each in TNT Buffer (or other validated wash buffer) at room temperature with agitation.
Step 9: Visualization of Deposited Fluorophores
For fluorescent detection, counterstain and mount for fluorescence microscopy. (See page 10 table, Fluorophore
Excitation and Emission Maxima.)
For chromogenic detection with TSA Fluorescein, incubate slides with anti-fluorescein-HRP (NEF710001EA) or antifluorescein-AP (NEF709001PK) followed by detection with an appropriate chromogenic substrate.

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Troubleshooting
Technical Support Resources• Email:• Telephoneo USA toll-freeo EU toll-freeo Finland toll-freeo China toll-free

global.techsupport@perkinelmer.com
800-762-4000
00800 33 29 0000
999 800 33 29 0000
800 820 5046

ISH Troubleshooting
REMEDY
PROBLEM
Low Signal

Excess Signal

High
Background

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Optimize probe concentration.
Titer HRP conjugate to determine optimum concentration for signal amplification.
Add tissue permeabilization step to facilitate penetration of reagents.
Lengthen incubation time for Fluorophore Tyramide Working Solution.
Consider TSA Plus reagents for higher signal.
Decrease concentration of HRP conjugate introduced prior to amplification.
Decrease probe concentration.
Decrease Fluorophore Tyramide Working Solution incubation time.
Decrease concentration of anti-fluorescein-enzyme conjugate used for chromogenic
visualization.
Decrease probe concentration.
Decrease concentration of HRP conjugate.
Check for endogenous biotin (if using streptavidin conjugates)
Shorten chromogenic development time.
Lengthen endogenous peroxidase quenching step.
Filter buffers.
Increase number and/or length of washes.
Nonqualified or contaminated blocking reagent used. Use PerkinElmer Blocking Reagent(FP1020 or FP1012).

IHC Troubleshooting
PROBLEM

REMEDY

Low Signal•

Excess Signal

High
Background

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Titer primary and/or secondary antibodies to determine optimum concentration for signal
amplification
Lengthen incubation time for Fluorophore Tyramide Working Solution.
Use antigen retrieval techniques to unmask the target.
Consider TSA Plus reagents for higher signal.
Decrease concentration of primary and/or secondary antibody or HRP conjugates.
Decrease Fluorophore Tyramide Working Solution incubation time.
Decrease concentration of anti-fluorescein-enzyme conjugate used for chromogenic
visualization..
Filter buffers
Decrease concentration of primary and/or secondary antibody or HRP conjugates.
Lengthen endogenous peroxidase quenching step.
Check for endogenous biotin (if using streptavidin conjugates)
Increase number and/or length of washes. Shorten chromogenic development time.
Nonqualified or contaminated blocking reagent used. Use PerkinElmer Blocking Reagent(FP1020 or FP1012).

Selected References
A quantitative evaluation of peroxidase inhibitors for tyramide signal amplification mediated cytochemistry andhistochemistry
G. Li, S. Amin, N. N. Okuhama, G. Liao, L. A. Mingle
Histochem Cell Biol (2006)
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1713225
Dual Fluorescent In Situ Hybridization and Immunohistochemical Detection with Tyramide Signal Amplification
Aliya U. Zaidi, Hideki Enomoto, Jeffrey Milbrandt, and Kevin A. RothJ Histochem Cytochem. (2000)
http://www.ncbi.nlm.nih.gov/pubmed/10990490
Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization
Tim Brend, Scott A. Holley J Vis Exp. (2009)http://www.ncbi.nlm.nih.gov/pubmed/19322135
Genomic Anatomy of the Hippocampus
Carol L. Thompson, Sayan D. Pathak, Andreas Jeromin, Lydia L. Ng, Cameron R. MacPherson, Marty T. Mortrud, Allison
Cusick, Zackery L. Riley, Susan M. Sunkin, Amy Bernard, Ralph B. Puchalski, Fred H. Gage, Allan R. Jones, Vladimir B.
Bajic, Michael J. Hawrylycz and Ed S. Lein
Neuron. (2008)
http://www.ncbi.nlm.nih.gov/sites/entrez/19109908?dopt=Abstract&holding=f1000,f1000m,isrctn
Detection of microRNAs in frozen tissue sections by fluorescence in situ hybridization using locked nucleic acidprobes and tyramide signal amplification
Asli N Silahtaroglu, Dorrit Nolting, Lars Dyrskjøt, Eugene Berezikov, Morten Møller, Niels Tommerup & Sakari Kauppinen
Nat Protoc. (2007)
http://www.ncbi.nlm.nih.gov/pubmed/17947994
Introduction of Tyramide Signal Amplification (TSA) to Pre-embedding Nanogold–Silver Staining at the Electron
Microscopic Level
Seung-won Lee, Song Eun Lee, Seong Hyuk Ko, Eun Kyoung Hong, Kwang Il Nam, Kei-ichiro Nakamura, Shuhei
Imayama, Yeong-Joon Park, Kyu Youn Ahn, Choon Sang Bae, Baik Yoon Kim, and Sung Sik ParkJ Histochem Cytochem. (2005)
http://www.ncbi.nlm.nih.gov/pubmed/15684338

Fluorophore Excitation and Emission Maxima
Fluorophore

Excitation

Emission

Coumarin

402 nm443 nmFluorescein

494 nm

517 nmTetramethylrhodamine

550 nm

570 nmCyanine 3

550 nm

570 nmCyanine 5

648 nm

667 nm

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Complementary Products
3g

FP1020

10 g

FP1012Anti‐rabbit IgG (goat) HRP

1 mL

NEF812001EA

Anti‐rabbit IgG (goat) HRP

100 µLNEF812E001EA

Anti‐mouse IgG (goat) HRP

1 mL

NEF822001EA

Anti‐mouse IgG (goat) HRP

100 µLNEF822E001EA

Anti‐human IgG (goat)* HRP

1 mL

NEF802001EA

Anti‐DNP‐HRP

150 µLFP1129Antifluorescein‐HRP

2x250µL

NEF710001EAStreptavidin‐HRP

2x250µL

NEL750001EA

Anti‐digoxigigenin HRP

500 µLNEF832001EA

Anti‐rabbit IgG (goat) biotin

1 mL

NEF813001EA

Anti‐mouse IgG (goat) biotin

1 mL

NEF823001EA

Anti‐human IgG (goat) biotin

1 mL

NEF803001EA

Anti‐digoxigigenin biotin

500 µLNEF833001EAStreptavidin Fluorescein

1 mL

NEL720001EAStreptavidin Texas Red®

1 mL

NEL721001EAStreptavidin Coumarin

1 mL

NEL722001EAStreptavidin‐HRP

2x250µL

NEL750001EAStreptavidin‐AP

2x250µL

NEL751001EATSA Blocking Reagent
Horseradish Peroxidase Reagents

Biotin Conjugates

Labeled Streptavidin

Chromogens
BCIP/NBT SubstrateDAB Substrate

For detection of Alkaline
Phosphatase
For detection of Horseradish
Peroxidase

NEL937001PK
NEL938001EA

Alkaline Phosphatase Reagents
Anti‐Mouse IgG (Goat), AP‐Labeled

1 mL

NEF824001EA

Anti‐Rabbit IgG (Goat), AP‐Labeled

1 mL

NEF814001EAStreptavidin‐ AP Conjugate

2x250µL

NEL751001EA

Antifluorescein‐AP Conjugate

2x250µL

NEF709001PK

Anti‐DNP‐AP

150 µLFP1131

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Hapten Labeled Deoxynucleotides (25 nmol, for labeling of ISH probes)
3‐Amino‐3‐Deoxydigoxigenin‐9‐dCTP

25 nmol

NEL562001EA

Biotin‐11‐dATP

25 nmol

NEL540001EA

Biotin‐11‐dCTP

25 nmol

NEL538001EA

Biotin‐11‐dGTP

25 nmol

NEL541001EA

Biotin‐11‐dUTP

25 nmol

NEL539001EA

DNP‐11‐dUTP

25 nmol

NEL551001EA

Fluorescein‐12‐dATP

25 nmol

NEL465001EA

Fluorescein‐12‐dCTP

25 nmol

NEL424001EA

Fluorescein‐12‐dGTP

25 nmol

NEL429001EA

Fluorescein‐12‐dUTP

25 nmol

NEL413001EA

Hapten Labeled Ribonucleotides (25 nmol, for labeling of ISH probes)
Biotin‐11‐ATP

250 nmol

NEL544001EA

Biotin‐11‐CTP

250 nmol

NEL542001EA

Biotin‐11‐GTP

250 nmol

NEL545001EA

Biotin‐11‐UTP

250 nmol

NEL543001EA

Fluorescein‐12‐ATP

250 nmol

NEL439001EA

Fluorescein‐12‐CTP

250 nmol

NEL434001EA

Fluorescein‐12‐GTP

250 nmol

NEL496001EA

Fluorescein‐12‐UTP

250 nmol

NEL414001EATSA Cyanine 3 and 5 reagents and related methods are covered by U.S. Patent 5,688,966 and international equivalents thereof.
For Research use only. This product is distributed and sold for research purposes only by the end-user in the research market, and, to that extent, by purchasing this productthe end-user is granted a limited license to use this product for research use only. This product is not intended for diagnostic or therapeutic use and no license or right isgranted for use of this product for diagnostic or therapeutic purposes. Purchase does not include or carry any right or license to use, develop or otherwise exploit this productcommercially. Any commercial use, development or exploitation of this product without the express prior written authorization of PerkinElmer is strictly prohibited and may
constitute infringement of the intellectual property rights of PerkinElmer under the aforementioned patents.
TSA is a registered trademark of PerkinElmer. Other trademarks are property of their respective owners.

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