Protein Electrophoresis and Western Blotting Education Center
We offer specialized kits for the detection of histidine-tagged proteins, post-translational modifications, multiple target proteins on a single western blot, and target proteins to verify siRNA reagent gene knockdown.
Fast western blotting
Pierce Fast Western Blotting Kits offer optimized reagents that shorten the time to perform a typical western blot from 4h down to approximately 55 mins. These kits help reduce hands-on and overall blotting time using Thermo Scientific ECL and SuperSignal Chemiluminescent substrates.
Each kit includes all reagents necessary to complete a western blot being probed with a mouse or rabbit primary antibody. The protocol requires minimal hands-on time and yields results comparable to classic western blotting with ECL.
| Thermo Scientific product | Sensitivity | Order |
|---|---|---|
| Fast Western Blot Kit, Pierce ECL | Picogram | Mouse and Rabbit (35050) |
| Fast Western Blot Kit, SuperSignal West Pico | Low pictogram | Mouse (35065) Rabbit (35066) |
| Fast Western Blot Kit, SuperSignal West Dura | Femtogram | Mouse (35075) Rabbit (35076) |
| Fast Western Blot Kit, SuperSignal West Femto | Low femtogram | Mouse (35085) Rabbit (35086) |
Tagged protein detection
We offer innovative kits for His-tagged and biotinylated protein detection. These are complete western blotting detection kits to detect the target protein in complex mixtures (cell lysates, tissues).
| Thermo Scientific product | Description | Order |
|---|---|---|
| SuperSignal West HisProbe Kit | Contains HisProbe-HRP Conjugate, as well as reagents required for detection of polyhistidine-tagged proteins by western blotting. HisProbe-HRP conjugate is a nickel (Ni2+)-activated derivative of horseradish peroxidase (HRP) that enables direct, IMAC-based detection of His-tagged proteins. | 15168 |
| Pierce Far-Western Blot Kit for Biotinylated Proteins | Enables in-gel or on-membrane western blot detection of target proteins via interaction with any biotin-labeled test protein used as the primary probe. The kit includes reagents and procedures for probing and chemiluminescent detection with blots or directly from mini gels. | 23500 |
Post translational modification detection
Label and detect S-nitrosylated proteins with this superior alternative to biotin switch assays for measuring the post-translational modification: S-nitrosylation.
| Thermo Scientific product | Description | Order |
|---|---|---|
| Pierce S-Nitrosylation Western Blot Kit | Enables sensitive detection of protein S-nitrosocysteine post-translational modifications in a complete, easy-to-use kit. | 90105 |
Biotin binding proteins as probes
The highly specific affinity interaction between biotin and avidin or streptavidin proteins can be used as a form of indirect detection using a primary or secondary antibody that is labeled with a biotin affinity tag. Then a secondary (or tertiary) probe, such as streptavidin that is labeled with a detectable enzyme or fluorophore tag, can be used to probe for the biotin tag to yield a detectable signal.
We offer streptavidin, neutravidin and avidin protein conjugates for biotinylated antibody or protein probe detection including HRP, AP conjugates and high-intensity fluorescent and infrared conjugates. Avidin, streptavidin, and neutravidin bind very strongly and specifically to biotin. However, each protein has their limitations in certain assays. Avidin, is glycosylated which may lead to nonspecific lectin binding in assays. Streptavidin contains an RYD motif, a bacterial recognition sequence, which can cause background binding with certain samples. Another alternative is to use neutravidin protein which is an exclusive, deglycosylated form of Avidin that avoids the drawbacks of both native avidin and streptavidin.
Enzyme-labeled Streptavidin and Avidin Conjugates
| Thermo Scientific product | Description | Order |
|---|---|---|
| Streptavidin-HRP | Streptavidin in a purified form conjugated to peroxidase for substrate-based detection. Recommended western blot concentration: 2 µg/mL | 21126 |
| High sensitivity streptavidin-HRP | Exclusive peroxidase-conjugated streptavidin biotin-binding protein that provides signal amplification like poly-HRP with exceptional storage stability | 21130 |
| Avidin-HRP | Native avidin conjugated horseradish peroxidase | 21123 |
| NeutrAvidin-HRP | NeutrAvidin protein-HRP is a specially-prepared form of avidin that decreases background in biotin-binding conjugated to peroxidase for substrate based detection. | 31001 |
| Streptavidin-AP | Streptavidin in a purified form, conjugated to alkaline phosphatase. Recommended western blot concentration: 2 µg/mL | 21324 |
| NeutrAvidin-AP | NeutrAvidin protein-HRP is a specially-prepared form of avidin that decreases background in biotin-binding conjugated to peroxidase for substrate based detection. | 31002 |
Fluorescent streptavidin conjugates
| Product | Excitation (nm) | Emission (nm) | Order |
|---|---|---|---|
| Alexa Fluor 488 streptavidin | 495 | 519 | S11223 (1 mg) S32354 (0.5 mL) |
| Alexa Fluor 555 streptavidin | 555 | 565 | S21381 (1 mg) S32355 (0.5 mL) |
| Alexa Fluor 647 streptavidin | 650 | 668 | S21374 (1 mg) S32357 (0.5 mL) |
| Alexa Fluor 680 streptavidin | 679 | 702 | S21378 (1 mg) S32358 (0.5 mL) |
| Alexa Fluor 750 streptavidin | 749 | 775 | S21384 (1 mg) |
Fast western blotting kits
Tagged protein detection
Enzyme-labeled streptavidin and avidin conjugates
Fluorescent conjugates
Fast western blotting
Pierce Fast Western Blotting Kits offer optimized reagents that shorten the time to perform a typical western blot from 4h down to approximately 55 mins. These kits help reduce hands-on and overall blotting time using Thermo Scientific ECL and SuperSignal Chemiluminescent substrates.
Each kit includes all reagents necessary to complete a western blot being probed with a mouse or rabbit primary antibody. The protocol requires minimal hands-on time and yields results comparable to classic western blotting with ECL.
| Thermo Scientific product | Sensitivity | Order |
|---|---|---|
| Fast Western Blot Kit, Pierce ECL | Picogram | Mouse and Rabbit (35050) |
| Fast Western Blot Kit, SuperSignal West Pico | Low pictogram | Mouse (35065) Rabbit (35066) |
| Fast Western Blot Kit, SuperSignal West Dura | Femtogram | Mouse (35075) Rabbit (35076) |
| Fast Western Blot Kit, SuperSignal West Femto | Low femtogram | Mouse (35085) Rabbit (35086) |
Tagged protein detection
We offer innovative kits for His-tagged and biotinylated protein detection. These are complete western blotting detection kits to detect the target protein in complex mixtures (cell lysates, tissues).
| Thermo Scientific product | Description | Order |
|---|---|---|
| SuperSignal West HisProbe Kit | Contains HisProbe-HRP Conjugate, as well as reagents required for detection of polyhistidine-tagged proteins by western blotting. HisProbe-HRP conjugate is a nickel (Ni2+)-activated derivative of horseradish peroxidase (HRP) that enables direct, IMAC-based detection of His-tagged proteins. | 15168 |
| Pierce Far-Western Blot Kit for Biotinylated Proteins | Enables in-gel or on-membrane western blot detection of target proteins via interaction with any biotin-labeled test protein used as the primary probe. The kit includes reagents and procedures for probing and chemiluminescent detection with blots or directly from mini gels. | 23500 |
Post translational modification detection
Label and detect S-nitrosylated proteins with this superior alternative to biotin switch assays for measuring the post-translational modification: S-nitrosylation.
| Thermo Scientific product | Description | Order |
|---|---|---|
| Pierce S-Nitrosylation Western Blot Kit | Enables sensitive detection of protein S-nitrosocysteine post-translational modifications in a complete, easy-to-use kit. | 90105 |
Biotin binding proteins as probes
The highly specific affinity interaction between biotin and avidin or streptavidin proteins can be used as a form of indirect detection using a primary or secondary antibody that is labeled with a biotin affinity tag. Then a secondary (or tertiary) probe, such as streptavidin that is labeled with a detectable enzyme or fluorophore tag, can be used to probe for the biotin tag to yield a detectable signal.
We offer streptavidin, neutravidin and avidin protein conjugates for biotinylated antibody or protein probe detection including HRP, AP conjugates and high-intensity fluorescent and infrared conjugates. Avidin, streptavidin, and neutravidin bind very strongly and specifically to biotin. However, each protein has their limitations in certain assays. Avidin, is glycosylated which may lead to nonspecific lectin binding in assays. Streptavidin contains an RYD motif, a bacterial recognition sequence, which can cause background binding with certain samples. Another alternative is to use neutravidin protein which is an exclusive, deglycosylated form of Avidin that avoids the drawbacks of both native avidin and streptavidin.
Enzyme-labeled Streptavidin and Avidin Conjugates
| Thermo Scientific product | Description | Order |
|---|---|---|
| Streptavidin-HRP | Streptavidin in a purified form conjugated to peroxidase for substrate-based detection. Recommended western blot concentration: 2 µg/mL | 21126 |
| High sensitivity streptavidin-HRP | Exclusive peroxidase-conjugated streptavidin biotin-binding protein that provides signal amplification like poly-HRP with exceptional storage stability | 21130 |
| Avidin-HRP | Native avidin conjugated horseradish peroxidase | 21123 |
| NeutrAvidin-HRP | NeutrAvidin protein-HRP is a specially-prepared form of avidin that decreases background in biotin-binding conjugated to peroxidase for substrate based detection. | 31001 |
| Streptavidin-AP | Streptavidin in a purified form, conjugated to alkaline phosphatase. Recommended western blot concentration: 2 µg/mL | 21324 |
| NeutrAvidin-AP | NeutrAvidin protein-HRP is a specially-prepared form of avidin that decreases background in biotin-binding conjugated to peroxidase for substrate based detection. | 31002 |
Fluorescent streptavidin conjugates
| Product | Excitation (nm) | Emission (nm) | Order |
|---|---|---|---|
| Alexa Fluor 488 streptavidin | 495 | 519 | S11223 (1 mg) S32354 (0.5 mL) |
| Alexa Fluor 555 streptavidin | 555 | 565 | S21381 (1 mg) S32355 (0.5 mL) |
| Alexa Fluor 647 streptavidin | 650 | 668 | S21374 (1 mg) S32357 (0.5 mL) |
| Alexa Fluor 680 streptavidin | 679 | 702 | S21378 (1 mg) S32358 (0.5 mL) |
| Alexa Fluor 750 streptavidin | 749 | 775 | S21384 (1 mg) |
Fast western blotting kits
Tagged protein detection
Enzyme-labeled streptavidin and avidin conjugates
Fluorescent conjugates
Resources
Download brochure: Western Detection Workflow
Explore: Pre-stained Protein Ladders
Explore: Western Blot Transfer Systems
For Research Use Only. Not for use in diagnostic procedures.