4-Hydroxynonenal 2D Western Blot
Our 4-Hydroxynonenal (4-HNE) Protein 2D Western Blot platform offers high sensitivity detection and accurate quantitation of 4-HNE modified Protein in any protein samples. 4-HNE, a highly toxic and most abundant end product of lipid peroxidation, has been implicated in the tissue damage, dysfunction and injury associated with aging and other pathological states such as cancer, Alzheimer, diabetes, cardiovascular and inflammatory complications. Further, 4-HNE, as a oxidative stress marker, plays a key role in cell signaling transduction. Applied Biomics’ 2D Fluorescent Western Blot has been successfully applied in studying the functions of 4-HNE modified Proteins.
Example 1: Mouse liver protein
4-HNE forms protein adducts during oxidative stress. This modification adversely affects the structure and functions of many proteins, and has been hypothesized to play a key role in cell signal transductions. In the following example, proteins from drug treated Mouse liver were extracted, followed by 4-HNE Protein 2D Western Blot using Applied Biomics’ protocols.
Mouse Liver Protein
Protein / Anti-4-HNE
Please follow these general principles regarding biohazardous materials and buffer condition in getting your samples ready.
Protein amount for each WB: 100-500 µg of total protein from each sample.
Protein concentration: 5-20 mg/ml is preferred.
Primary antibody requirement for each WB: Sufficient for 100ml incubation volume, and need information on:
- Animal species in which the antibody was raised
- Suggested dilution factor
If you cannot provide the dilution factor, please let us know:
- If it is a commercial antibody, provide the link to the antibody information
- If it is not a commercial antibody, please provide the protein concentration of the antibody, whether it is IgG fraction or anti-sera or antigen-affinity purified.
Buffer: Please feel free to submit samples in whichever buffer that you would normally use. The standard sample preparation is covered by the DIGE gel cost. Such samples should have protein concentration in the range of 5-20 mg/ml. Additional charge may apply on samples that require extensive amount of the extra work such as protein eluting, concentrating, buffer exchange or serum abundant protein depletion.
Sample type: Please inform our scientists about your sample type. We will send you additional tips for each sample type.
|2D Gel with 1 CyDye Labeling 1
|2D DIGE Gel with 2 CyDye Labeling 1
|Transfer to PVDF / Nitrocellulose Membrane 2
|Western Blot 1-color 3
|Western Blot 2-color 3
|Call for price
|Call for price
Primary antibody: Customers are responsible for providing the primary antibodies. Applied Biomics will provide the following primary antibodies at $150-250 for each Western blot:
- Anti-Bacteria HCP antibody: E. coli; Lactococcus lactis
- Anti-Mammalian Cell HCP antibody: Vero Cell; CHO; BHK; HEK 293; A549; MRC5; PER.C6
- Anti-Yeast HCP antibody: Saccharomyces cerevisiae; Pichia pastoris
- Anti-Insect Cell HCP antibody: SF9; SF22
- Others: Anti-Bovine Serum Protein antibody; Anti-Mycobacterium Protein antibody; Anti-Goat-Milk Protein antibody
Fluorescent labeled secondary antibody: Provided by Applied Biomics and included in the price.
1. Price covers:
- Experimental design
- Standard sample preparation and protein concentration determination
- Fluorescent dye labeling using CyDye
- IEF and SDS-PAGE
- Gel image scan using Typhoon scanner
- Image analysis (ImageQuant)
- Data Report
2. Price covers:
- Trans-blotting 2D Gel to PVDF or Nitrocellulose membrane
- Membrane images scanned after the transfer
3. Price covers:
- Western blot using the primary antibodies provided by customers, Applied Biomics will provide secondary antibodies
- Membrane images scanned after Western Blot
1-3. Each Western blot will cover 2 hour data analysis generating Protein and Western blot image report.
Price does NOT cover:
- Primary antibody provided by customer
- Manual matching spots and manual circling and numbering spots on power point images
- Customized data request in addition to the standard request