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Simplify Your Western: Six Simple Steps


Western blotting is viewed as the gold standard for protein detection in molecular biology research. It is used to identify proteins within a cell or tissue lysate. Antibodies against your protein(s) of interest, bind to specific epitopes to identify the target protein within a lysate. Due to the high specificity of the binding, multiple target proteins can be identified on one membrane. Secondary antibodies then bind to your primary antibodies and when exposed to a substrate react allowing for the visualization of the corresponding protein band.


Western Blotting Process Step 1
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Who wants to spend all that time in the dark?


We have removed the need for you to spend time in the dark room.
Let your blot develop before your eyes, right there on your bench!




No need to spend time in the dark room

Signal Develops In Front Of Your Eyes

The WESTERNVIEW? Detection Kits for Western Blot analysis quickly and clearly reveal bands on the transfer membrane without the need for specialized equipment.


The most common method of visualization still seen today is via a chemiluminescent reaction. This involves spending time in a tight dark room, with a timer and a list of exposure times, which are mostly chosen based on educated guesswork.

  • A simple solution for real-time results – no film developer or fluorescent reader required or time in the dark room.
  • High sensitivity – quickly visualize your bands with minimal overexposure risk and with no visible background.
  • Convenient and easy to use – includes secondary antibody and all reagents necessary to visualize your bands.

For high sensitivity, ease of use, and long-lasting signal, try our new WESTERNVIEW Detection Kits



Product Kit Contents
WESTERNVIEW Detection Kit (Anti-Mouse) AP Anti-Mouse Secondary, NBT/BCIP, Antibody Diluent, 10x Wash Buffer
WESTERNVIEW Detection Kit (Anti-Rabbit) AP Anti-Rabbit Secondary, NBT/BCIP, Antibody Diluent, 10x Wash Buffer
WESTERNVIEW Dual Detection Kit (Anti-Mouse / Anti-Rabbit) HRP Anti-Mouse Secondary, AP Anti-Rabbit Secondary, NBT/BCIP, DAB Chromogen, DAB Substrate Buffer, Antibody Diluent, SignaSure? Butter Salts, Tween?-20



Reliably Distinguish Multiple Targets


PANC-1 cells treated with or without etoposide. Loaded total cell lysate (20 μg) and simultaneously incubated with tubulin and caspase antibodies before being developed with the WESTERNVIEW Dual Detection Kit (Anti-Mouse / Anti-Rabbit).
Multiple Target Western Blot





Sensitive Results Without Expensive Equipment


Total cell lysate from Jurkat cells. Membrane was probed with anti-tubulin. Detected using WESTERNVIEW Detection Kit (Anti-Mouse) and exposed for 1 minute.
Western Blot sensitivity



Clear Signals Without the Background


Jurkat cells treated with or without 12.5 μM etoposide for 18 hours. Loaded 12 μg lysate per well.
Strong Western Blot Signal

Western Blot E-book

  • Western Blot: Principle and Theory
  • What are the Differences Between Northern, Southern, and Western Blotting?
  • Ten Tips for Successful Westerns
  • Five Western Blot Problems and How to Troubleshoot Them
  • How Do You Choose the Right Western Blot Detection Method?
  • Membrane Selection: A Quick Comparison of PVDF and Nitrocellulose

Primary Antibody for Western Blotting

We have over 2,000 antibodies validated for Western Blot.

  • Specific – to bind your protein of interest or primary antibody
  • Sensitive – for high detection and low background
  • Reliable – for consistent results
  • Validated – for Western blot application and stated species

Primary Antibody for Western Blot Search Tool



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Antibodies for Loading Controls


Find the right loading control to ensure proper interpretation of your Western blots. The sample type will help you determine which target to use for your loading control. Typically, loading controls are housekeeping proteins expressed at high and consistent levels.

Loading controls are used to:
  • Check for equal transfer across membrane
  • Ensure even loading between lanes
  • Normalize and quantify your data



Sample Type Target MW (kDa) Product Name
Whole Cell & Cytoskeleton Vinculin 124 Vinculin (human) monoclonal antibody (FB11)
Alpha Tubulin 50 α-Tubulin Recombinant monoclonal antibody (F2C) (Mouse IgG1λ)
α-Tubulin Recombinant monoclonal antibody (F2C) (Rabbit IgGλ)
Beta Tubulin 50 β-Tubulin Recombinant monoclonal antibody (S11B) (Mouse IgG1λ)
β-Tubulin Recombinant monoclonal antibody (S11B) (Rabbit IgGλ)
Actin 42 Actin polyclonal antibody
Actin clonal antibody (S12-I)
Beta Actin 42 β-Actin polyclonal antibody
GAPDH 36 GAPDH monoclonal antibody (1D4)
GAPDH monoclonal antibody (GA1R) (HRP conjugate)
Mitochondrial HSP60 60 HSP60 monoclonal antibody (LK-1)
HSP60 monoclonal antibody (LK-2)
HSP60 monoclonal antibody (Mab11-13)
HSP60 polyclonal antibody
VDAC1 31 Voltage-dependent anion channel polyclonal antibody
Nuclear Lamin B1 66 Laminin B chain (human) monoclonal antibody (DG10)
HDAC1 5 HDAC1 polyclonal antibody
PCNA 29 PCNA monoclonal antibody (SPM350)
Serum Transferrin 77 Transferrin polyclonal antibody

Blotting Membranes

Optimized for Western Transfer - Polyvinylidene Fluoride (PVDF) Membranes


  • Sensitive protein detection with low background and very low burn-through
  • Provides high surface area for strong hydrophobic interactions (adsorb 50% more protein than nylon or nitrocellulose.)
  • Broad solvent compatibility


Membrane Specifications
Protein Binding Capacity 100-300 μg/cm2
Solvent Resistant Resistant to acetone, DMSO, dimethyl formahide, methanol, trifluoroacetic acid, and triethylamine.
Binding Interaction Hydrophobic
Pore Size 0.2 μM
Total Protein Stain Compatibility Amido black, Ponceau S, Colloidal gold, Colloidal silver, India ink, Coomassie blue
Double-blotting Method Yes
Strip and Re-probe Yes
Detection Methods Chromogenic, Chemiluminescent, Fluorescent, Radioactive, Chemifluorescent
Other Applications Amino Acid Analysis, Protein Sequencing, Solid Phase Assay Systems
Available Formats PVDF Transfer Membrane (7 x 9 cm)
PVDF Transfer Membrane (10 X 15 cm)


Prestained Protein Ladder

Protein ladder for use with protein gel electrophoresis

  • 3 μL or 5 μL per loading for clear visualization during electrophoresis on 15-well or 10-well mini-gel, respectively
  • Three color protein standard with 13 prestained proteins
  • Apply more for thicker (>1.5 mm) or larger gel

Our Protein ladder is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins. The ladder is supplied in gel loading buffer and is ready to use. Do not heat, dilute or add reducing agent before loading.

Proteins are covalently coupled with a blue chromophore except for two reference bands (one green and one red band at 25 kDa and 75 kDa respectively) when separated on SDS-PAGE (Trisglycine buffer).


Prestained Protein Ladder for Western Blot

Guide for Molecular Weight Estimation (kDa): Migration patterns of Prestained Protein Ladder in different electrophoresis conditions.
 

Blocking Solutions

Product Size Kit Contents
BSA Solution 50 mL 10% BSA in PBS

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