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Avidin DCS, Texas Red® labeled, is a cell sorter grade of Texas Red® Avidin D. Through the development of a new method of labeling we are able to incorporate more fluorochrome into a protein without altering the properties of the protein. Not only does the Texas Red® Avidin DCS have a high F/P ratio, the efficiency of fluorescence is also increased. In addition to the improved conjugation technique, we employ a purification step to reduce nonspecific binding which can hamper resolution in cell sorting and other applications.
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Unit Size
1 mg
Applications
Immunofluorescence, In situ hybridization, Flow Cytometry/Cell Separation
Recommended Usage
For diluting this product, we recommend a HEPES- or bicarbonate-buffered saline solution, approximately pH 8.2. The recommended range of concentration for staining of cells or tissues is 5-20 µg/ml.
Solution
10 mM HEPES, 0.5 M NaCl, pH 7.5, 0.08% sodium azide, 10 mg/ml beta cyclodextrin
VECTASHIELD® Antifade Mounting Medium Catalog #: H-1000-10
VECTASHIELD® Antifade Mounting Medium with DAPI Catalog #: H-1200-10
VECTASHIELD® HardSet™ Antifade Mounting Medium Catalog #: H-1400-10
VECTASHIELD® HardSet™ Antifade Mounting Medium with DAPI Catalog #: H-1500-10
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Although this product has been designed for cell sorting, it can also be used in fluorescent microscopy when higher sensitivity and lower background staining are necessary. This derivative is preferred when using the amplification procedure with Biotinylated Anti-Avidin D.
Excitation maximum: 595 nm
Emission maximum: 615 nm
Color: Red
Home All Products Fluorophore Conjugated Avidin/Streptavidin Texas Red® Avidin DCS, Cell Sorting Grade
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AMCA Streptavidin is produced by conjugating streptavidin with a coumarin fluorescent dye, 7-amino-4-methylcoumarin-3-acetic acid. This derivative excites in the ultraviolet (350 nm) and emits in the visible (450 nm) producing an intense blue fluorescence.
Amplification of fluorescent signals can be easily achieved with our biotinylated secondary antibodies followed by our highly purified fluorochrome-labeled streptavidin or avidin. Using a biotin/avidin or biotin/streptavidin detection system results in an additional layer of amplification over a directly conjugated secondary antibody.
Features:
Recommended for routine immunofluorescence applications
Highly purified and possesses very low non-specific binding properties
Extremely high affinity for biotin
Has a high fluorochrome to protein ratio
Compared to conventional primary and secondary fluorescent techniques, can provide greater sensitivity and lower background staining
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More Information
Unit Size
1 mg
Applications
Immunofluorescence, In situ hybridization, Flow Cytometry/Cell Separation
Recommended Usage
For diluting this product, we recommend a HEPES- or biocarbonate-buffered saline solution, approximately pH 8.2.Avoid using RPMI 1640 or other biotin-containing solutions as diluents.Serum also can contain biotin and should not be added to diluents. The recommended concentration range for use is 10-30 µg/ml.
Solution
10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.08% sodium azide
VECTASHIELD® Antifade Mounting Medium Catalog #: H-1000-10
VECTASHIELD® Antifade Mounting Medium with Propidium Iodide (PI) Catalog #: H-1300-10
VECTASHIELD® HardSet™ Antifade Mounting Medium Catalog #: H-1400-10
VECTASHIELD® HardSet™ Antifade Mounting Medium with Phalloidin Catalog #: H-1600-10
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Vector Laboratories fluorochrome-conjugated streptavidin and avidin reagents are highly purified and possess very low non-specific binding properties. They have extremely high affinity for biotin. These fluorescent conjugates can be used to detect biotinylated secondary antibodies and other macromolecules in applications such as immunofluorescence, in situ hybridization, or flow cytometry.
Most paraffin-embedded tissues have little autofluorescence under those conditions. AMCA Streptavidin may provide an advantage in many labeling situations over conjugates of fluorescein and related dyes. AMCA Streptavidin can be used on frozen or paraffin-embedded sections as a single label or in combination with other fluorescent conjugates for multiple labeling studies.
Home All Products Fluorophore Conjugated Avidin/Streptavidin Streptavidin, AMCA
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The QuantTag Biotin Kit (BDK-2000) is designed to determine the amount of free biotin in solution or the number of biotin molecules attached to nucleic acids, proteins, or other macromolecules. This kit can be used to accurately determine the labeling efficiency of your process to label molecules with biotin.
The kit contains enough material to perform 25 assays using the 1 ml protocol (including the creation of a standard curve for each) or 250 assays using the 0.1 ml protocol.
*Note: Avoid contact with skin and eyes. If contact occurs, flush immediately with water.
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User Guide
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Related Products
Biotin (Long Arm) Hydrazide Catalog #: SP-1100-50
Biotin (Long Arm) NHS Catalog #: SP-1200-50
Biotin (Long Arm) NHS, Water Soluble Catalog #: SP-1210-50
PHOTOPROBE® Biotin for Nucleic Acid Labeling Catalog #: SP-1000
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The QuantTag Biotin Kit (BDK-2000) is designed to determine the amount of free biotin in solution or the number of biotins attached to nucleic acids, proteins, or other macromolecules. This kit can be used to accurately determine the labeling efficiency of your process.
Unlike conventional biotin quantitation methods like the HABA assay, no predigestion of protein or nucleic acids is required, which saves time and increases accuracy. The QuantTag Biotin Kit, being more sensitive than the HABA assay, is able to detect less than 1 nmol of biotin. QuantTag Kit reagents chemically react with free or bound biotin producing a colored product that can be quantified using a spectrophotometer. The absorbance is measured in the visible spectrum allowing the use of plastic cuvettes or microtitre plates.
The QuantTag Biotin Kit is quick and easy to use and the assay can be completed in 30 minutes. The kit contains sufficient material to perform from 25 to 250 tests depending on assay size. A biotin standard is included.
Home All Products Biotin and Digoxigenin Labeling Reagents QuantTag Biotin Quantitation Kit
I recently purchased Normal Goat Serum, Cat. No. S-1000, from Vector Laboratories for use as a general block in my tissue staining applications. What is the percentage of serum, and once diluted, how long can I store the working solution?
Normal Goat Serum, Cat. No. S-1000, is essentially spun whole blood, filtered, heat inactivated and preserved with the addition of sodium azide. The 20 ml provided therefore is all goat serum. For most IHC and IF tissue staining applications, an aliquot is taken and diluted in assay buffer such as PBS or TBS to 2-10% (v/v). This working solution can be stored in the fridge and used for up to one week. Beyond this time, there would be concern for possible microbial contamination that would affect performance.
Related Products
Normal Horse Serum Blocking Solution Catalog #: S-2000-20
Normal Horse Serum Blocking Solution, 2.5% Catalog #: S-2012-50
Normal Chicken Serum Blocking Solution Catalog #: S-3000-20
Normal Swine Serum Blocking Solution Catalog #: S-4000-20
Normal Rabbit Serum Blocking Solution Catalog #: S-5000-20
Normal Goat Serum Blocking Solution Catalog #: S-1000-20
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Home All Products Protein Blocking Solutions Normal Goat Serum Blocking Solution, 2.5%
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The ImmPRESS polymerized reporter enzyme staining system uses novel conjugation and micropolymer chemistries to create a highly sensitive, ready-to-use, one-step, non-biotin detection system for immunohistochemistry and immunocytochemistry staining. This unique micropolymer of highly active horseradish peroxidase (HRP) is attached to our affinity purified secondary antibodies, producing reagents with outstanding sensitivity and low background.
Features:
Enhanced accessibility to nuclear and membrane antigens
Ready-to-use in a convenient dropper bottle
Shorter assay time
Simplified multiple labeling
Designed to detect mouse primary antibodies on rat tissues that may contain endogenous rat immunoglobulins
The ImmPRESS (15 ml) Kit will stain approximately 75-150 sections.
Specifications
More Information
Unit Size
15 ml
Applications
Immunohistochemistry / Immunocytochemistry, In situ hybridization, ELISAs
Technology
Micropolymer Reagents
Detection Enzyme(s)
Peroxidase
Conjugate
Micropolymer HRP
Target Species
Mouse
Format
Ready-to-Use
Kit Components
Kit Contents:
15 ml ImmPRESS (Peroxidase) Polymer Anti-Mouse IgG, Rat Adsorbed Reagent (made in horse, ready-to-use)
15 ml 2.5% Normal Horse Serum for blocking (ready-to-use)
Documents
ImmPRESS Polymer & Automated Staining App. Note
ImmPRESS Troubleshooting Guide
User Guide
Safety Data Sheet
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What are the advantages of using ImmPRESS® Excel kits compared with one-step peroxidase polymer detection systems?
The ImmPRESS Excel kits are a two-step peroxidase polymer based detection system that include an unconjugated amplifier antibody. This intermediate amplifier antibody increases sensitivity of the assay at least three- to four-fold over that of a one-step polymer based system, by facilitating the introduction of more peroxidase enzyme at the site of specific antigen localization. This increase in sensitivity would be advantageous in instances of weak antigen expression, and to further dilute out an expensive primary antibody. The ImmPRESS Excel Amplifier kits are presented as a complete kit format that include enzyme quench, blocking serum and ImmPACT® DAB EqV peroxidase substrate, in addition to the amplifier antibody and defined ImmPRESS polymer secondary antibody. However, as the two published references below indicate, this format is still modular and allows for the substitution of different detection reagents. These references describe using ImmPACT NovaRED™ peroxidase substrate in combination with the ImmPRESS Excel Amplifier kits. Oncogenesis (2017) 6, e293; doi:10.1038/oncsis.2016.82 Tan, E.M.S., et al (2017) Front. Med. 4:162 (doi: 10.3389/fmed.2017.00162)
Our research lab has an open automated staining platform. Do you know if the ImmPRESS reagents can be applied to an automated system, and if so, do you have any guidelines or procedures?
Yes, we conducted studies on the compatibility of the ImmPRESS HRP polymer reagents with three commercially available autostainers (Agilent/Dako Autostainer Plus, Leica Bond Rx, and Ventana Discovery Ultra). We showed that our reagents are suitable for IHC detection on each of these platforms. The ImmPRESS polymer reagents and enzyme substrates generated equivalent IHC staining results compared to reagents from the instrument manufacturer. Some modifications in the protocol were performed to optimize the signal-to-noise ratio, such as a shorter incubation time for the ImmPRESS polymer and increase in the number of buffer washes following polymer incubation. Please see the following application note for more details about each automated platform:https://info.vectorlabs.com/l/360741/2019-01-11/2hv9mw
Related Products
ImmPACT® DAB Substrate, Peroxidase (HRP) Catalog #: SK-4105
Normal Horse Serum Blocking Solution, 2.5% Catalog #: S-2012-50
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The peroxidase micropolymers of the ImmPRESS HRP polymer reagent limit steric interference and provide enhanced accessibility to the target, avoiding the disadvantages of other polymer systems that use large dextrans or other macromolecules as backbones. The result is crisp, strong staining of antibody targets, especially nuclear and membrane antigens (such as Ki67, estrogen receptor, bcl-2, CD3, CD8 and CD10) and greater sensitivity than other polymer systems.
Staining Procedure
The staining procedure is simple as shown in the diagram below. Following a blocking step with the diluted normal horse serum, sections are incubated with primary antibody. After a brief wash, the appropriate ImmPRESS Reagent is added to the sections and incubated for 30 minutes. Sections are again rinsed and the slides are developed with the peroxidase substrate of choice.
Consider Species Cross-Reactivity
When choosing the optimal detection system for your application, it is important to consider not only the species of the primary antibody but also the species of the tissue. If the species of the primary antibody and the species of the tissue are closely related (e.g. rat and mouse), the secondary antibody may bind to endogenous IgG in the tissue section leading to background. The following options minimize background staining in these instances:
Use a secondary antibody specifically adsorbed to remove cross-reacting antibodies of closely-related species (e.g. ImmPRESS Anti-Mouse IgG, Rat Adsorbed).
Use the M.O.M. ImmPRESS Kit (Cat. No. MP-2400) for applications of mouse primary antibodies on mouse tissue.
Learn more about the ImmPRESS Polymer Detection Systems in double labeling.
Product review on Biocompare.com.
Home All Products ImmPRESS Peroxidase (HRP) Polymer Reagents ImmPRESS® HRP Horse Anti-Mouse IgG, Rat adsorbed Polymer Detection Kit, Peroxidase
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The ImmPACT VIP HRP substrate produces an intense, violet (purple) reaction product, and can be used as an alternative to DAB or as a second color for multiple antigen labeling. This product generates an intense reaction product that is 2-4 times more sensitive than the Vector VIP substrate kit (SK-4600).
Features:
2-4 fold greater sensitivity than original substrate kit
Optimized for IHC/ICC/ISH and blots
Permanent mounting
Ideal for single and multiple labeling
Working solutions stable for up to 14 days at 4 °C
Supplied as 120 ml of diluent and stock solutions in dropper bottles
Specifications
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Unit Size
120 ml
Applications
Immunohistochemistry / Immunocytochemistry, In situ hybridization, Blotting Applications, Elispot
VectaMount® Permanent Mounting Medium Catalog #: H-5000-60
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Vector VIP and ImmPACT VIP chromogens also provide excellent color contrast in pigmented tissues such as melanoma or retina (1). Sections stained with either substrate can be viewed by darkfield and electron microscopy. Vector VIP and ImmPACT VIP substrates can be used for both manual and automated staining methods. Stained sections should be dehydrated, cleared, and permanently mounted.
This kit consists of 120 ml of diluent and concentrated stock solutions of ImmPACT VIP reagents in convenient dropper bottles. Working solutions are stable for up to 14 days at 4 °C.
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The Vector SG HRP substrate produces a blue-gray reaction product that can be used as a single label or as a second color for multiple antigen labeling.
Features:
Greater sensitivity than conventional substrates
Consistent and reliable
Ideal for IHC/ICC/ISH and blots
Permanent mounting
No wait times for mixing and dissolving powders or tablets
Specifications
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Unit Size
1 kit
Applications
Immunohistochemistry / Immunocytochemistry, In situ hybridization, Blotting Applications, Elispot
Nuclear Fast Red Counterstain Catalog #: H-3403-500
Methyl Green Counterstain Catalog #: H-3402-500
VectaMount® Permanent Mounting Medium Catalog #: H-5000-60
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Sections stained with the substrate also can be viewed by darkfield or electron microscopy. Vector SG and ImmPACT SG substrates can be used for both manual and automated staining methods. Stained sections can be dehydrated, cleared, and permanently mounted.
Vector SG Substrate Kit contains stock solutions in convenient dropper bottles. The kit provides all of the necessary reagents to prepare about 300 ml of working solution.
Home All Products Horseradish Peroxidase (HRP) Substrates Vector® SG Substrate Kit, Peroxidase (HRP)
VectaMount® Permanent Mounting Medium Catalog #: H-5000-60
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Vector NovaRED and ImmPACT® NovaRED HRP substrates produce red reaction products. Unlike AEC, sections stained with either substrate should be dehydrated, cleared, and permanently mounted. Both chromogens are useful as alternatives to DAB or as a second color for multiple antigen labeling. Vector NovaRED and ImmPACT NovaRED substrates also provide excellent color contrast in pigmented tissue such as melanoma. With the aid of imaging systems and software, the spectral profile of both substrates can be distinguished from other enzyme substrates in applications where antigens are co-localized. Sections stained with either substrate also can be viewed by darkfield microscopy. Vector NovaRED and ImmPACT NovaRED substrates can be used for both manual and automated staining methods.
Vector NovaRED Substrate Kit contains stock solutions in convenient dropper bottles. The sensitivity of Vector NovaRED substrate is equivalent to Vector DAB Substrate, and 4 times greater than AEC. This product can also be used on blots. This kit provides all of the necessary reagents to prepare about 300 ml of working solution. When stored at 4 °C, this kit is stable for one year.
Home All Products Horseradish Peroxidase (HRP) Substrates Vector® NovaRED® Substrate Kit, Peroxidase (HRP)
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Vector Methyl Green stains cell nuclei light green. It is an excellent alternative in multiple labeling when hematoxylin obscures the substrate colors.
Features:
Simple, two-step procedure
Suitable for use with a wide range of enzyme substrates
Suitable for use with non-aqueous mountants
Specifications
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Unit Size
500 ml
Applications
Immunohistochemistry / Immunocytochemistry, In situ hybridization
Format
Ready-to-Use
Documents
User Guide
Counterstain/Substrate Compatibility Chart
Safety Data Sheet
Download CoA
Datasheet
Product FAQs
I am using Methyl Green (H-3402) as a counterstain for my IHC, but the stain is a little weak. What suggestions do you have to increase the staining intensity?
Methyl Green counterstain does take a little bit of optimization in some applications. We would suggest heating a volume of the counterstain (~300 ml) to 60 °C and add to a Coplin jar or similar glass staining rack, and submerse the slides. This approach facilitates a better uptake of stain into the section compared with placing the slide on a heated surface and placing drops of stain onto the section. Following the incubation time (~3-5 min), remove slides and wash in tap water. Omit the acetone, acetic acid rinse step described in the instructions and move the slides directly into 95% ethanol for the dehydration, clearing and mounting process. This methodology will retain more stain in the nuclei and hence produce greater intensity.
Related Products
Hematoxylin QS Counterstain Catalog #: H-3404-100
VectaMount® Permanent Mounting Medium Catalog #: H-5000-60
Nuclear Fast Red Counterstain Catalog #: H-3403-500
Hematoxylin Counterstain Catalog #: H-3401-500
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Counterstain/Substrate Compatibility Table
This table is designed as a reference to determine the optimal counterstain/substrate combination for your application.
Home All Products Counterstains and Special Stains Methyl Green Counterstain
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