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Topic: Western blot


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In the News (Wed 21 Aug 19)

  
  Reference.com/Encyclopedia/Western blot
A western blot (alternately, immunoblot) is a method to detect a specific protein in a given sample of tissue homogenate or extract.
The name western blot was given to the technique by W. Neal Burnette and is a play on the name Southern blot, a technique for DNA detection developed earlier by Edwin Southern.
The colorimetric detection method depends on incubation of the western blot with a substrate that reacts with the reporter enzyme (such as peroxidase) that is bound to the secondary antibody.
www.reference.com /browse/wiki/Western_blot   (2637 words)

  
  Western Blot by Teresa Burnette
In molecular biology, blotting means the transfer of macromolecules both nucleic acids and proteins, from one solid or semisolid gel to another usually from a gel to a filter paper.
The practical uses of the western blotting test include diagnosing infection, screening blood units to reduce the risk of HIV transmission, and testing the immunogenicity and protective efficacy of a new vaccine.
Western blot analysis measures for the presence of antibodies to several HIV proteins.
www.samford.edu /~gekeller/burnette.html   (626 words)

  
 Interpretation and Use of the Western Blot Assay for Serodiagnosis of Human Immunodeficiency Virus Type 1 Infections
The Western blot assay is a method in which individual proteins of an HIV-1 lysate are separated according to size by polyacrylamide gel electrophoresis.
The criteria for a negative Western blot interpretation specify "no bands." This interpretation is essential because some observed bands may reflect the presence of antibodies to HIV regulatory proteins or may indicate partially processed or degraded viral structural proteins.
However, when a Western blot test has only the multimeric form of gp41 and no true gp120 present, a serum sample would be scored as positive on the basis of the presence of antibody to a single envelope glycoprotein, gp41.
www.cdc.gov /MMWR/preview/mmwrhtml/00001431.htm   (2579 words)

  
 Western Blot Unreliability
Such stringent guidelines require the Western blot to be an extremely precise test, capable of definitively identifying the exact antibodies that show up in a patient's serum sample.
In theory, according to Fawcett, the Western blot linearly separates all known proteins of the LD spirochete, Borrelia burgdorferi (Bb).
His study, "Use of Western blot and Enzyme-Linked Immunosorbent Assays in the Diagnosis of Lyme Disease" (Pediatrics, 1991:88:465-470) is cited in the study that became the basis for the CDC criteria.
www.lyme.org /lymelight/westernblot.html   (1107 words)

  
 Western Blot
In molecular biology, a Western blot is a method used to detect proteins in a tissue sample or extract.
Like the related Southern and Northern blots, it separates denatured proteins with gel electrophoresis and then transfers the proteins onto a membrane with a blotting technique.
The Western blot is primarily used in medical diagnostic applications like testing for mad-cow disease and some forms of Lyme disease.
www.iscid.org /encyclopedia/Western_Blot   (219 words)

  
 Western Blot Detection System
Western immunoblotting is a highly sensitive technique which relies on the specificity of antibodies to identify proteins that have been electrophoretically transferred from SDS-Polyacrylamide gels to solid support membranes.
Western blot: 10 µg samples of rat brain lysate (lanes 1, 2, 3) were resolved by electrophoresis, transferred to nitrocellulose and probed using the antibodies described below.
Lane 3: Stripped blot in lane 2 was reprobed using an anti-ERK2 polyclonal antibody at 1µg/mL; secondary antibody, goat antirabbit IgG, HRP conjugate (AP132P), 1:2,500.
www.chemicon.com /featured/westernblot.asp   (332 words)

  
 The Western Blot Test
The Western Blot test places multiple proteins (antigens) believed to be from HIV in a line on a test strip (according to molecular weight).
Since the Western Blot is an antibody test, like the initial ELISA test, it is not fully independent, and the very common use of this as a confirmatory test for ELISA is thus questionable.
on commercial Western blot strips, are primarily multimers of the HIV transmembrane glycoprotein gp41 that react with antibodies to gp41.
www.rethinkaids.com /quotes/test-wb.html   (1904 words)

  
 Western Blot
Western blotting can give you information about the size of your protein (with comparison to a size marker or ladder in kDa), and also give you information on protein expression (with comparison to a control such as untreated sample or another cell type or tissue).
Western blot is dependent on the quality of antibody you use to probe for your protein of interest, and how specific it is for this protein.
Western blot analysis examines proteins, and so we want the proteins to be release from the cells and also prevent the proteins from being cut up by proteases.
www.molecularstation.com /protein/western-blot   (1943 words)

  
 Virtual Cell Labs: Transfer Step
In a Western Blot, however, electrophoresis is used as a way to transfer proteins from one medium to another instead of pulling them through a medium to separate them.
To run the blot, final assembly of the electrophoresis apparatus is necessary, and then the voltage is set and applied for a period of time.
Specifically, the interior of the blot module is filled with transfer buffer, and the exterior area is filled with water or transfer buffer.
www.bme.gatech.edu /vcl/WesternBlotting/Background/transfer.htm   (789 words)

  
 Western blot analysis with quantum dot fluorescence technology: a sensitive and quantitative method for multiplexed ...
Western or immunoblotting analysis of protein expression in cells and tissues has been the major analytical tool for assessing molecular biological functions in basic cell biological research, pathology and drug discovery and development.
For example, a blot with serial dilutions of the p42 mitogen-activated phosphokinase (p42 MAPK) protein from 5 ng to 20 pg per lane was imaged for 5, 2 and 0.6 min (Fig.
Graph of mean band intensity of recombinant p42 MAPK from western blot images acquired at (a) 0.6 min and (b) 2 min exposure shown in Figure 1.
www.nature.com /nmeth/journal/v2/n1/full/nmeth0105-79.html   (1652 words)

  
 C. Brenner: Explanation of the Lyme Disease Western Blot
First of all, it should be noted that the Western blot is usually performed as a follow-up to an ELISA test, which is the most commonly employed initial test for Lyme disease.
Both the ELISA and the Western blot are "indirect" tests -- that is, they measure the immune system's response to an infectious agent rather than looking for components of the agent itself.
Finally, it is important to keep in mind that no matter how carefully the Western blot test is carried out and interpreted, its usefulness, like that of all tests that measure B. burgdorferi antibodies, is ultimately contingent on the reliability of the human immune response as an indicator of exposure to B. burgdorferi.
www.lymenet.de /labtests/brenner.htm   (2530 words)

  
 Genotech - Western Blot Analysis
One such widely used method is known as Western Blot analysis.
Western Blot analysis involves electrophoresis separation of test protein samples on polyacrylamide gels then the separated protein samples are electroblotted on a protein binding membrane – where protein is captured and immobilized.
In this lab activity, students carry out Western Blot procedure and screen test samples from a population of infected individuals and group them according to the precise nature of their disease.
www.gbiosciences.com /Western-Blot-desc.aspx   (138 words)

  
 westernblotting.org - western blot protocols, troubleshooting, electrophoresis and science information
In western blotting (or immunoblotting), prior to protein immobilization on the PVDF or nitrocellulose membranes, sample proteins are separated using SDS polyacrylamide gel electrophoresis (SDS-PAGE) providing information about molecular weight and the potential existence of different isoforms of the proteins under study.
Western blotting provides a method for protein expression level comparisons, however several factors need to be considered.
"Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem.
www.westernblotting.org   (1043 words)

  
 Chemicon - Product #2060 - ReBlot™ Western Blot Recycling Kit
Western blotting is a commonly used technique for studying protein function and localization.
The CHEMICON Re-Blot Western Blot Recycling Kit is effective for removal of antibodies from Western blots that have been developed with chemiluminescence or radioactive iodine or other isotopes.
The Re-Blot Western Blot Recycling Kit should be used only for qualitative purposes until it has been established by comparative blot analysis that stripping does not quantitatively affect a given antigen.
www.chemicon.com /browse/productdetail.asp?ProductID=2060   (600 words)

  
 Western Blot Test - Questions and Answers
Both the IHC and the Western blot are recognized by the World Animal Health Organization as confirmatory tests for BSE and can be used under the U.S. BSE surveillance protocol.
The Western blot test is the only other means of confirmatory testing authorized under the USDA protocol, which are consistent with the OIE standards for BSE surveillance.
A. The IHC and Western blot are considered to be equally specific; they simply use different means to test for the disease.
www.mad-cow-facts.com /News-Commentary/western-blot-test-05.htm   (1158 words)

  
 [No title]
Unlike the use in donor counseling, if a licensed Western blot has been performed as the additional, more specific test for reentry purposes, additional IFA testing may not be used to negate or resolve a positive or indeterminate Western blot result.
For example, if a licensed western blot test result is indeterminate, whether on the initial, substituted, or follow-up sample, a negative IFA result obtained subsequently is not sufficient to qualify a donor for reentry.
The results of additional testing on the donor sample by either IFA or Western blot may be provided to the consignees of such products as a basis for the decision to trace and notify recipients of previously collected units from currently positive donors.
www.fda.gov /Cber/bldmem/042392.txt   (970 words)

  
 aidsmap.com | Western Blot
The Western Blot test is a kind of ELISA test that distinguishes between antibodies that recognise different antigens in the test system.
Western Blot relies on a subjective judgement of the intensity of the bands of antibody which form in response to particular antigens.
Western Blot should NOT be used as a stand-alone HIV antibody test, but only as part of a multi-test strategy.
www.aidsmap.com /cms1008317.asp   (588 words)

  
 Visualizer™ Western Blot Detection Kits
The Visualizer™ Western Blot Detection Kit was designed to make Western detection of proteins easier by providing the highest level of sensitivity with the least amount of background in a chemiluminescent reagent system.
Upstate's Visualizer™ Western Blot Detection Kit utilizes a superior version of the chemiluminescent HRP substrate luminol that results in the fastest and most sensitive detection of an antigen while providing a long lasting signal and the highest signal to noise ratio of any competing product on the market.
Blots were blocked for 1 hour at room temperature in 5% nonfat milk PBST.
www.komabiotech.co.kr /product/electrophoresis/visualizer.htm   (701 words)

  
 Western blotting
Western blot analysis can detect one protein in a mixture of any number of proteins while giving you information about the size of the protein.
Western blotting tells you how much protein has accumulated in cells.
Also, if a protein is degraded quickly, Western blotting won't detect it well; you'll need to use (RIP).
lifesciences.asu.edu /resources/mamajis/western/western.html   (829 words)

  
 ZeptoMetrix Corp-WESTERN BLOTS
The ZeptoMetrix SIV Western Blot assay is a qualitative enzyme immunoassay for the in vitro detection of antibodies to SIV in serum or plasma.
HELICO BLOT 2.1 Western blot kit is a qualitative assay for the detection of IgG antibodies to Helicobacter pylori (H.
HIV-1 BLOT 1.3 is a qualitative enzyme immunoassay for the detection of antibodies to HIV-1 in human serum or plasma.
www.zeptometrix.com /blots.htm   (370 words)

  
 Western Blot Protocol
Transfer the proteins to a nitrocellulose membrane (SandS NCTM) at 250 mA in transfer buffer for 1-4 h, depending on the size of the target protein.
Incubate the blot with primary antibody (diluted 1:250 to 1:1000 in blocking buffer) for 1 h in blocking buffer at RT.
Incubate blot with anti-rabbit IgG-HRP conjugate (Sigma) (diluted 1:10,000 - 1:2,000 in blocking buffer) for 1 h in blocking buffer at RT.
www.prosci-inc.com /Antibody/Western-Blot-Procedure.html   (171 words)

  
 Pierce Biotechnology - Restore Western Blot Stripping Buffer
The same blot was stripped wtih Restore Western Blot Stripping Buffer (B) for 5 minutes at room temperature and re-probed (C) with the primary antibody at 1/5,000 and the HRP-secondary conjugate at 1:20,000.
Nitrocellulose Western blots of EGF-stimulated A431 cell lysate proteins (4, 2 and 1 µg in lanes 1, 2 and 3, respectively) were detected with Pierce ECL Substrate (Pierce #32209).
Blot B was stripped using 5 ml of Restore PLUS Western Blot Stripping Buffer for 15 minutes at room temperature (Panel 2) and was re-probed for non-phosphorylated form of MAPK½ using anti-MAP Kinase ½ (Erk ½) rabbit polyclonal (Upstate) at 0.5 µg/ml and goat anti-rabbit-HRP (Product # 31460, Blot 3B) at 200 ng/ml.
www.piercenet.com /Objects/View.cfm?type=ProductFamily&ID=02040210   (1198 words)

  
 Visualizer(tm) Western Blot Detection Kit
Upstate's Visualizer™ Western Blot Detection Kit utilizes a superior version of the chemiluminescent HRP substrate luminol that results in the fast and extremely sensitive levels of detection of an antigen while providing a long lasting signal with signal-to-noise ratios as high or higher than any competing product on the market.
Each blot was incubated in the detection reagent as listed and laid side-by-side for 30 seconds exposure to film.
Legend: Western blot — SDS-PAGE gel with a whole adult rat brain lysate, resolved by electrophoresis, transferred to a PVDF membrane and probed for target protein.
www.upstate.com /features/visualizer.asp   (1050 words)

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