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).

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.

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 Localization of creatine kinase isoenzymes in myofibrils. II. Chicken heart muscle.

1977 ◽  
Vol 75 (2) ◽  
pp. 318-325 ◽  
Author(s):  
T Wallimann ◽  
H J Kuhn ◽  
G Pelloni ◽  
D C Turner ◽  
H M Eppenberger
Keyword(s):  
Skeletal Muscle ◽  
Creatine Kinase ◽  
Heart Muscle ◽  
Fluorescent Antibody ◽  
Immunofluorescent Staining ◽  
Absolute Amount ◽  
Chicken Heart ◽  
Antibody Staining ◽  
Line Region ◽  
Almost All

Chicken heart muscle contains almost exclusively the BB isoenzyme of creatine kinase (CK), its myofibrils, moreover, lack an M-line. This tissue thus provides an interesting contrast to skeletal muscle, in which some of the MM-CK present as predominant CK isoenzyme is bound at the myofibrillar M-line. Approx. 2% of the total CK activity in a chicken heart homogenate remains bound to the myofibrillar fraction after repeated washing cycles; both the fraction and the absolute amount of CK bound are about threefold lower than in skeletal muscle. Almost all of the bound enzyme is located within the Z-line region of each sarcomere, as revealed by indirect fluorescent-antibody staining with antiserum against purified chicken BB-CK. After incubation with exogenous purified MM-CK, positive immunofluorescent staining for M-type CK at the H-region of heart myofibrils was observed, along with weaker fluorescence in the Z-line region. Chicken heart myofibrils may thus possess binding sites for both M and B forms of CK.

scholarly journals Sarcomere-Length Dependence of the Low Angle X-Ray Pattern from Skeletal Muscle Fibers at Rest and during Isometric Contraction

2012 ◽  
Vol 102 (3) ◽  
pp. 147a-148a
Author(s):  
Gabriella Piazzesi ◽  
Massimo Reconditi ◽  
Elisabetta Brunello ◽  
Luca Fusi ◽  
Marco Linari ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Isometric Contraction ◽  
Sarcomere Length ◽  
Muscle Fibers ◽  
X Ray ◽  
Length Dependence ◽  
Skeletal Muscle Fibers
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