scholarly journals THE M BAND

1971 ◽  
Vol 48 (2) ◽  
pp. 340-347 ◽  
Author(s):  
Ellen Kundrat ◽  
Frank A. Pepe
Keyword(s):  
Strong Evidence ◽  
Structural Integrity ◽  
Staining Pattern ◽  
Fluorescent Antibody ◽  
Extraction Procedure ◽  
Thick Filament ◽  
Tris Buffer ◽  
Antibody Staining ◽  
Band Material ◽  
Fluorescent Antibody Staining

The M band can be extracted from fibrils suspended in 5 mM Tris buffer, pH 8.0, for 15 min. The M band is completely removed only from fibrils of sarcomere lengths greater than 2.1 µ. Extraction does not alter the fluorescent antimyosin staining pattern of the A band, thus providing strong evidence that no alteration of the structural integrity of the thick filament has occurred. Fluorescent antibody staining of the M band of unextracted fibrils can be prevented specifically by absorbing the fluorescent antibody with extracted M band material prior to staining. This verifies the specificity of the extraction procedure.

scholarly journals Immunofluorescent and histochemical localization of AMP deaminase in skeletal muscle.

1979 ◽  
Vol 81 (2) ◽  
pp. 361-373 ◽  
Author(s):  
B Ashby ◽  
C Frieden ◽  
R Bischoff
Keyword(s):  
Skeletal Muscle ◽  
Sarcomere Length ◽  
Staining Pattern ◽  
Fluorescent Antibody ◽  
Muscle Fibers ◽  
Histochemical Localization ◽  
Amp Deaminase ◽  
Heavy Meromyosin ◽  
Antibody Staining ◽  
Fluorescent Antibody Staining

Fluorescent antibody staining experiments with both isolated myofibrils and muscle fibers grown in culture show that AMP deaminase is bound to the myofibril in the A band. The strongest staining occurs at each end of the A band. The approximate width of the fluorescent stripes and their relation to the A band remains constant as a function of sarcomere length. Removal of enzyme from the myofibrils leads to loss of staining, and readdition of purified enzyme restores the original staining pattern. A histoenzymatic method for the detection of AMP deaminase activity in cultured fibers gives comparable localization. The results are consistent with the previous observation (Ashby, B. and C. Frieden. 1977.J. Biol. Chem. 252:1869--1872) that AMP deaminase forms a tight complex in solution with subfragment-2 (S-2) of myosin or with heavy meromyosin (HMM).

Profilin is predominantly associated with monomeric actin in Acanthamoeba

1999 ◽  
Vol 112 (21) ◽  
pp. 3779-3790 ◽  
Author(s):  
D.A. Kaiser ◽  
V.K. Vinson ◽  
D.B. Murphy ◽  
T.D. Pollard
Keyword(s):  
Monoclonal Antibodies ◽  
Plasma Membrane ◽  
Fluorescent Antibody ◽  
Gel Filtration ◽  
Live Cells ◽  
A431 Cells ◽  
Antibody Staining ◽  
Large Particles ◽  
Biochemical Fractionation ◽  
Fluorescent Antibody Staining

We used biochemical fractionation, immunoassays and microscopy of live and fixed Acanthamoeba to determine how much profilin is bound to its known ligands: actin, membrane PIP(2), Arp2/3 complex and polyproline sequences. Virtually all profilin is soluble after gentle homogenization of cells. During gel filtration of extracts on Sephadex G75, approximately 60% of profilin chromatographs with monomeric actin, 40% is free and none voids with Arp2/3 complex or other large particles. Selective monoclonal antibodies confirm that most of the profilin is bound to actin: 65% in extract immunoadsorption assays and 74–89% by fluorescent antibody staining. Other than monomeric actin, no major profilin ligands are detected in crude extracts. Profilin-II labeled with rhodamine on cysteine at position 58 retains its affinity for actin, PIP(2) and poly-L-proline. When syringe-loaded into live cells, it distributes throughout the cytoplasm, is excluded from membrane-bounded organelles, and concentrates in lamellapodia and sites of endocytosis but not directly on the plasma membrane. Some profilin fluorescence appears punctate, but since no particulate profilin is detected biochemically, these spots may be soluble profilin between organelles that exclude profilin. The distribution of profilin in fixed human A431 cells is similar to that in amoebas. Our results show that the major pool of polymerizable actin monomers is complexed with profilin and spread throughout the cytoplasm.

scholarly journals Detection ofCryptosporidium molnariOocysts from Fish by Fluorescent-Antibody Staining Assays forCryptosporidiumspp. Affecting Humans

2011 ◽  
Vol 77 (5) ◽  
pp. 1878-1880 ◽  
Author(s):  
Rona Barugahare ◽  
Michelle M. Dennis ◽  
Joy A. Becker ◽  
Jan Šlapeta
Keyword(s):  
Ribosomal Dna ◽  
Fluorescent Antibody ◽  
Small Subunit ◽  
Rapid Diagnosis ◽  
Antibody Staining ◽  
Direct Fluorescent Antibody ◽  
Murray Cod ◽  
Diagnosis Of Infection ◽  
Formalin Fixed ◽  
Fluorescent Antibody Staining

ABSTRACTThree direct fluorescent-antibody staining assay kits for the detection of zoonoticCryptosporidiumspecies were used to detectCryptosporidium molnarifrom Murray cod, and the cryptosporidia were characterized by using small-subunit (SSU) ribosomal DNA (rDNA). To facilitate rapid diagnosis of infection, this study demonstrated that all three kits detected freshC. molnariand two kits detected formalin-fixed oocysts.

scholarly journals Quantitative immunohistochemistry: a comparison of microdensitometric analysis of unlabeled antibody peroxidase-antiperoxidase staining and of microfluorometric analysis of indirect fluorescent antibody staining for nicotinamide adenosine dinucleotide phosphate (NADPH)-cytochrome c (P-450) reductase in rat liver.

1983 ◽  
Vol 31 (10) ◽  
pp. 1183-1189 ◽  
Author(s):  
M T Smith ◽  
J A Redick ◽  
J Baron
Keyword(s):  
Cytochrome C ◽  
Rat Liver ◽  
Immunohistochemical Staining ◽  
Fluorescent Antibody ◽  
Indirect Fluorescent Antibody ◽  
Liver Lobule ◽  
Nadph Cytochrome ◽  
Antibody Staining ◽  
Unlabeled Antibody ◽  
Fluorescent Antibody Staining

The intralobular distribution of nicotinamide adenine dinucleotide phosphate (NADPH)-cytochrome c (P-450) reductase (NADPH:ferricytochrome oxidoreductase, EC 1.6.2.4) in rat liver has been investigated by means of two quantitative immunohistochemical techniques: microdensitometric quantitation of unlabeled antibody peroxidase-antiperoxidase staining and microfluorometric analysis of indirect fluorescent antibody staining. Utilizing sheep antiserum elicited against NADPH-cytochrome c (P-450) reductase that had been isolated and purified to apparent homogeneity from rat liver microsomes, the reductase was detected within hepatocytes throughout the liver. However, differences in the intensity of staining of hepatocytes within different regions of the liver lobule were readily apparent after completion of both immunohistochemical staining procedures. These visual findings were verified by microdensitometric and microfluorometric analyses of immunohistochemical staining, both of which revealed that approximately the same degree of staining for NADPH-cytochrome c (P-450) reductase was produced within the centrilobular and midzonal regions of the liver lobule, whereas periportal hepatocytes were stained with significantly less intensity. These results demonstrate that the application of either microdensitometry in conjunction with unlabeled antibody peroxidase-antiperoxidase staining or microfluorometry after indirect fluorescent antibody staining can be used to quantitatively determine the intratissue distributions of antigens.

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