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Detection of Blotted Proteins

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Cover of 'Detection of Blotted Proteins'

Table of Contents

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    Book Overview
  2. Altmetric Badge
    Chapter 1 Western Blotting: Origin and Ascent of the Species.
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    Chapter 2 Methods to Concentrate Proteins for Protein Isolation, Proteomic, and Peptidomic Evaluation.
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    Chapter 3 Measuring Protein Concentration on Nitrocellulose and After the Electrophoretic Transfer of Protein to Nitrocellulose
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    Chapter 4 Detection of Blotted Proteins: Not All Blockers Are Created Equal
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    Chapter 5 Protein Stains to Detect Antigen on Membranes
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    Chapter 6 Fluorescent Labeling of Proteins and Its Application to SDS-PAGE and Western Blotting
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    Chapter 7 Rapid, Antibody-Free Detection of Recombinant Proteins on Blots Using Enzyme Fragment Complementation
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    Chapter 8 Use of Nonradioactive Detection Method for North- and South-Western Blot
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    Chapter 9 Immunoblotting Using Radiolabeled Reagents for Detection
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    Chapter 10 Immunoblotting of Antigens: Whole, Strip, and New-Line Nitrocellulose Membrane Immunoblotting Using the Chemiluminescence Technique
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    Chapter 11 Detection of Protein Carbonyls by Means of Biotin Hydrazide–Streptavidin Affinity Methods
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    Chapter 12 Direct Immunodetection of Antigens Within the Precast Polyacrylamide Gel
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    Chapter 13 Quantitative Analysis of Signal Transduction with In-Cell Western Immunofluorescence Assays
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    Chapter 14 Ultrasensitive Protein Detection on Dot Blots and Western Blots with Semiconducting Polymer Dots
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    Chapter 15 Co-detection of Target and Total Protein by CyDye Labeling and Fluorescent ECL Plex Immunoblotting in a Standard Proteomics Workflow.
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    Chapter 16 Using Biotinylated Proteins to Demonstrate Immunodetection of Antigens via Western Blotting, Dot Blots, and Immunohistochemistry
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    Chapter 17 Calcium Binding by Ro 60 Multiple Antigenic Peptides on PVDF Membrane
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    Chapter 18 Sequential Use of Immunoblots for Characterization of Autoantibody Specificities
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    Chapter 19 Nanogold Immunodetection Detection Systems for the Identification of Autoantigens by Western Blotting
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    Chapter 20 Application of Intermittent Microwave Irradiation to Western Blot Analysis
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    Chapter 21 Visualization of Unstained Protein Bands on PVDF
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    Chapter 22 Multiplexed Fluorescent Immunodetection Using Low Autofluorescence Immobilon ® -FL Membrane
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    Chapter 23 Cold Microwave-Enabled Protein Detection and Quantification
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    Chapter 24 TLC-Blot (Far-Eastern Blot) and Its Application to Functional Lipidomics
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    Chapter 25 Analysis of Electroblotted Proteins by Mass Spectrometry.
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    Chapter 26 On-Membrane Renaturation of Recombinant Ro60 Autoantigen by Calcium Ions
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    Chapter 27 Phosphoprotein Detection on Protein Electroblot Using a Phosphate-Specific Fluorophore
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    Chapter 28 Purification of Tryptic Digests on Polyvinylidene Difluoride Membrane
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    Chapter 29 Detection of Blotted Proteins on Nitrocellulose/PVDF Membranes by Alta
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    Chapter 30 Nonstripping “Rainbow” and Multiple Antigen Detection (MAD) Western Blotting
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    Chapter 31 Supported Molecular Matrix Electrophoresis
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    Chapter 32 Parafilm-M(®), An Available Cost-Effective Alternative for Immuno-blot Pouches.
  34. Altmetric Badge
    Chapter 33 Succinylation-Alcian Blue Staining of Mucins on Polyvinylidene Difluoride Membranes
  35. Altmetric Badge
    Chapter 34 Comparison of Chemiluminescence vs. Infrared Techniques for Detection of Fetuin-A in Saliva
  36. Altmetric Badge
    Chapter 35 A Novel Methodology for Stripping and Reprobing of Western Blots Originally Developed with Colorimetric Substrate TMB
  37. Altmetric Badge
    Chapter 36 Other Notable Methods of Membrane Protein Detection: A Brief Review
  38. Altmetric Badge
    Chapter 37 Nitrocellulose Membrane: The New Canvas
  39. Altmetric Badge
    Chapter 38 Invisible Ink Marking in ECL Membrane Assays.
Attention for Chapter 32: Parafilm-M(®), An Available Cost-Effective Alternative for Immuno-blot Pouches.
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About this Attention Score

  • Above-average Attention Score compared to outputs of the same age (51st percentile)
  • Good Attention Score compared to outputs of the same age and source (73rd percentile)

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Chapter title
Parafilm-M(®), An Available Cost-Effective Alternative for Immuno-blot Pouches.
Chapter number 32
Book title
Detection of Blotted Proteins
Published in
Methods in molecular biology, January 2015
DOI 10.1007/978-1-4939-2718-0_32
Pubmed ID
Book ISBNs
978-1-4939-2717-3, 978-1-4939-2718-0
Authors

Quadri, Syed M S, Syed M. S. Quadri, Quadri, Syed M. S.

Abstract

Commercially available standard immuno-blot pouches do play an efficient role in antibody incubation in performing an immuno-blot, but are not readily available in the laboratory and have to be specifically ordered. We have developed an equally efficient technique to make an immune-blot more cost-effective with more conservation of antibodies by using a common and readily available laboratory product Parafilm-M(®). Parafilm-M(®) which serves as a sealant for various items of laboratory equipment can be used for antibody incubation. Manually made Parafilm-M(®) pouch has a clear advantage over standard immuno-blot pouches in terms of availability, cost-effectiveness, and consumption of antibodies that ultimately reduces the cost of an immuno-blot. We have performed a series of experiments to check the efficacy of both the techniques. Samples with equal amount of protein were analyzed on separate SDS PAGE gels. The proteins were transferred electrophoretically to the nitrocellulose membrane using Trans-Blot(®) Turbo™ Mini Nitrocellulose Transfer Pack. Antibody incubation was done using standard immuno-blot pouch, standard container and Parafilm-M(®) sealed pouch. The expression of protein was determined and the results of immuno-blots were compared. We found that antibodies are binding the membrane in Parafilm-M(®) pouches as efficiently as in container method or in standard immuno-blot pouches. By restricting the membrane, the surface area of the manually made Parafilm-M(®) pouch can be reduced, less diluent is required to cover the membrane as a result less antibodies are consumed. We also calculated that each immuno-blot pouch cost around $0.1906, whereas the cost for Parafilm-M(®) pouch is 0.0695 which is almost one-third the price of an immuno-blot pouch. Thus, Parafilm-M(®) method distinctly provides a cost-effective solution for antibody incubation.

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Attention Score in Context

This research output has an Altmetric Attention Score of 2. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 13 August 2020.
All research outputs
#9,520,538
of 16,190,116 outputs
Outputs from Methods in molecular biology
#2,608
of 9,357 outputs
Outputs of similar age
#157,628
of 345,935 outputs
Outputs of similar age from Methods in molecular biology
#286
of 1,189 outputs
Altmetric has tracked 16,190,116 research outputs across all sources so far. This one is in the 39th percentile – i.e., 39% of other outputs scored the same or lower than it.
So far Altmetric has tracked 9,357 research outputs from this source. They receive a mean Attention Score of 2.7. This one has gotten more attention than average, scoring higher than 69% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 345,935 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 51% of its contemporaries.
We're also able to compare this research output to 1,189 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 73% of its contemporaries.