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Detection System Kit Product

The Phototope-HRP Western Blot Detection System is designed for the chemiluminescent detection of proteins in standard Western blotting applications. Proteins and biotinylated molecular weight markers (provided) are separated by SDS-PAGE and transferred onto membrane. Following incubation with your primary anti-serum, horseradish peroxidase (HRP) linked secondary antibody and HRP-linked anti-biotin antibody are bound and then allowed to react with LumiGLO® reagent. The light emitted by destabilized LumiGLO® reagent is subsequently captured on X-ray film.

Background: Chemiluminescence systems have emerged as the best all-around method for western blot detection. They eliminate the hazards associated with radioactive materials and toxic chromogenic substrates. The speed and sensitivity of these methods are unequalled by traditional alternatives, and because results are generated on film, it is possible to record and store data permanently. Blots detected with chemiluminescent methods are easily stripped for subsequent reprobing with additional antibodies. HRP-conjugated secondary antibodies are utilized in conjunction with specific chemiluminescent substrates to generate the light signal. HRP conjugates have a very high turnover rate, yielding good sensitivity with short reaction times.

The Phototope-HRP Western Blot Detection System is designed for the chemiluminescent detection of proteins in standard Western blotting applications. Proteins and biotinylated molecular weight markers (provided) are separated by SDS-PAGE and transferred onto membrane. Following incubation with your primary anti-serum, horseradish peroxidase (HRP) linked secondary antibody and HRP-linked anti-biotin antibody are bound and then allowed to react with LumiGLO® reagent. The light emitted by destabilized LumiGLO® reagent is subsequently captured on X-ray film.

Background: Chemiluminescence systems have emerged as the best all-around method for western blot detection. They eliminate the hazards associated with radioactive materials and toxic chromogenic substrates. The speed and sensitivity of these methods are unequalled by traditional alternatives, and because results are generated on film, it is possible to record and store data permanently. Blots detected with chemiluminescent methods are easily stripped for subsequent reprobing with additional antibodies. HRP-conjugated secondary antibodies are utilized in conjunction with specific chemiluminescent substrates to generate the light signal. HRP conjugates have a very high turnover rate, yielding good sensitivity with short reaction times.

The BCA Protein Assay Kit can be used to measure the protein concentration of lysates or homogenates, in microplate format, prepared with the following buffers: Cell Lysis Buffer (10X) #9803, RIPA Buffer (10X) #9806, PathScan® Sandwich ELISA Lysis Buffer (1X) #7018. The dynamic range for this assay is 0.125 - 2 mg/mL. It is recommended that the BCA Compatibility Reagent be used to decrease interference from reducing agents, chelators, detergents, and other common ingredients found in most lysis buffers. Please see the attached protocol for additional details.
$42
120 slides
1 Kit
$140
1200 slides
1 Kit
The SignalStain® DAB Substrate Kit contains all of the necessary reagents to prepare a working solution of diaminobenzidine (DAB) for staining tissue sections. The DAB working solution reacts with peroxidase (HRP) detection systems such as the SignalStain® Boost IHC Detection Reagents (HRP, Rabbit #8114, and HRP, Mouse #8125), yielding a brown reaction product.
APPLICATIONS

Application Methods: Immunohistochemistry (Paraffin)

$30
25 ml each substrate
50 ml
$259
250 ml each substrate
500 ml
SignalFire™ ECL Reagent from Cell Signaling Technology (CST) is an enhanced chemiluminescent substrate capable of detecting picogram amounts of protein by western blot analysis. Compared to entry-substrates, SignalFire™ ECL Reagent boasts a more robust signal and extended duration of signal output. In the presence of hydrogen peroxide, horseradish peroxidase (HRP) converts luminol to an excited intermediate dianion. This dianion emits light on return to its ground state. Light emission is maximal immediately after exposure of the substrate to HRP and continues for 1 hour. Light can be captured on X-ray film, typically by exposure for a few seconds. Maximum sensitivity can be obtained with longer exposure.
APPLICATIONS

Application Methods: Western Blotting

Background: Chemiluminescence systems have emerged as the best all-around method for western blot detection. They eliminate the hazards associated with radioactive materials and toxic chromogenic substrates. The speed and sensitivity of these methods are unequalled by traditional alternatives, and because results are generated on film, it is possible to record and store data permanently. Blots detected with chemiluminescent methods are easily stripped for subsequent reprobing with additional antibodies. HRP-conjugated secondary antibodies are utilized in conjunction with specific chemiluminescent substrates to generate the light signal. HRP conjugates have a very high turnover rate, yielding good sensitivity with short reaction times.

$64
5 ml each
$205
25 ml each
LumiGLO®* chemiluminescent substrate is a luminol-based system designed for use with our Phototope®-HRP detection assays utilizing peroxidase-labeled antibodies immobilized on membranes. In the presence of hydrogen peroxide, horseradish peroxidase (HRP) converts luminol to an excited intermediate dianion. This dianion emits light on return to its ground state. Light emission is maximal immediately after exposure of the substrate to HRP and continues for 0.5-1 hour. Light can be captured on X-ray film, typically by exposure for a few seconds. Maximum sensitivity can be obtained by longer exposure. *Avoid repeated exposure to skin (see enclosed Material Safety Data Sheet or refer to our website for further information).
APPLICATIONS

Application Methods: Western Blotting

Background: Chemiluminescence systems have emerged as the best all-around method for western blot detection. They eliminate the hazards associated with radioactive materials and toxic chromogenic substrates. The speed and sensitivity of these methods are unequalled by traditional alternatives, and because results are generated on film, it is possible to record and store data permanently. Blots detected with chemiluminescent methods are easily stripped for subsequent reprobing with additional antibodies. HRP-conjugated secondary antibodies are utilized in conjunction with specific chemiluminescent substrates to generate the light signal. HRP conjugates have a very high turnover rate, yielding good sensitivity with short reaction times.

$81
10 ml each substrate
20 ml
$363
50 ml each substrate
100 ml
SignalFire™ Plus ECL Reagent from Cell Signaling Technology (CST) is a highly sensitive chemiluminescent substrate capable of detecting low picogram amounts of protein by western blot analysis. SignalFire™ Plus ECL Reagent has an extended duration of signal output lasting several hours following blot exposure, allowing for multiple exposures with either film or a digital imaging system. The strong signal output allows detection of low abundance proteins, conservation of reagents, and short exposure times.SignalFire™ Plus ECL Reagent requires approximately five-fold less Anti-rabbit IgG, HRP-linked Antibody #7074 or Anti-mouse IgG, HRP-linked Antibody #7076 than traditional ECL reagents. Limiting the amount of HRP exposed to the membrane prevents high background, oversaturation of the target protein signal, or false negative results. Other HRP-conjugated antibodies, including HRP-conjugated primary and anti-biotin-HRP antibodies, should be diluted similarly. Dilution of secondary antibody from alternative vendors may need to be optimized. Titration of lysate and primary antibody concentration is recommended to achieve optimal signal-to-noise ratio.
APPLICATIONS

Application Methods: Western Blotting

Background: Chemiluminescence systems have emerged as the best all-around method for western blot detection. They eliminate the hazards associated with radioactive materials and toxic chromogenic substrates. The speed and sensitivity of these methods are unequalled by traditional alternatives, and because results are generated on film, it is possible to record and store data permanently. Blots detected with chemiluminescent methods are easily stripped for subsequent reprobing with additional antibodies. HRP-conjugated secondary antibodies are utilized in conjunction with specific chemiluminescent substrates to generate the light signal. HRP conjugates have a very high turnover rate, yielding good sensitivity with short reaction times.

$86
10 ml each substrate
20 ml
$390
50 ml each substrate
100 ml
SignalFire™ Elite ECL Reagent from Cell Signaling Technology (CST) is an ultra sensitive chemiluminescent substrate capable of detecting femtogram amounts of protein by western blot analysis. SignalFire™ Elite ECL Reagent is compatible with both film and digital imaging systems. The extremely intense signal output allows detection of very low abundance proteins, conservation of reagents, and short exposure times.SignalFire™ Elite ECL Reagent requires approximately ten-fold less Anti-rabbit IgG, HRP-linked Antibody #7074 or Anti-mouse IgG, HRP-linked Antibody #7076 than traditional ECL reagents. Limiting the amount of HRP exposed to the membrane prevents high background, oversaturation of the target protein signal, or false negative results. Other HRP-conjugated antibodies, including HRP-conjugated primary and anti-biotin-HRP antibodies, should be diluted similarly. Dilution of secondary antibody from alternative vendors may need to be optimized. Titration of lysate and primary antibody concentration is recommended to achieve optimal signal-to-noise ratio.
APPLICATIONS

Application Methods: Western Blotting

Background: Chemiluminescence systems have emerged as the best all-around method for western blot detection. They eliminate the hazards associated with radioactive materials and toxic chromogenic substrates. The speed and sensitivity of these methods are unequalled by traditional alternatives, and because results are generated on film, it is possible to record and store data permanently. Blots detected with chemiluminescent methods are easily stripped for subsequent reprobing with additional antibodies. HRP-conjugated secondary antibodies are utilized in conjunction with specific chemiluminescent substrates to generate the light signal. HRP conjugates have a very high turnover rate, yielding good sensitivity with short reaction times.