scholarly journals Polyclonal Antisera Purification

2020 ◽  
Author(s):  
Keyword(s):  
Polyclonal Antisera

Use of monoclonal and polyclonal antibodies to detect prostatic acid phosphatase by immunoblotting.

1986 ◽  
Vol 32 (10) ◽  
pp. 1832-1835 ◽  
Author(s):  
P C Patel ◽  
L Aubin ◽  
J Côte
Keyword(s):  
Monoclonal Antibodies ◽  
Acid Phosphatase ◽  
Western Blot ◽  
Polyclonal Antibodies ◽  
Prostatic Acid Phosphatase ◽  
The Other ◽  
Dot Blot ◽  
Western Blots ◽  
Dot Blot Assay ◽  
Polyclonal Antisera

Abstract We investigated two techniques of immunoblotting--the Western blot and the dot blot--for use in detecting prostatic acid phosphatase (PAP, EC 3.1.3.2). We used polyclonal antisera to human PAP, produced in rabbits by hyperimmunization with purified PAP, and PAP-specific monoclonal antibodies in the immunoenzymatic protocols. We conclude that PAP can be readily detected by Western blots with use of polyclonal antisera, but not with monoclonal antibodies. On the other hand, using a dot blot assay, we could easily detect PAP with both polyclonal and monoclonal antibodies.

scholarly journals Evidence for Unknown Sarcocystis-Like Infection in Stranded Striped Dolphins (Stenella coeruleoalba) from the Ligurian Sea, Italy

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1201
Author(s):  
Federica Giorda ◽  
Umberto Romani-Cremaschi ◽  
Antoinette E. Marsh ◽  
Carla Grattarola ◽  
Barbara Iulini ◽  
...  
Keyword(s):  
Neospora Caninum ◽  
Positive Staining ◽  
Ligurian Sea ◽  
Sarcocystis Neurona ◽  
Stenella Coeruleoalba ◽  
Polyclonal Antisera ◽  
Ligurian Coast ◽  
Sarcocystis Falcatula ◽  
The Brain ◽  
The Mediterranean Basin

Two striped dolphins (SD1, SD2), stranded along the Ligurian coast of Italy, were diagnosed with a nonsuppurative meningoencephalitis associated with previously undescribed protozoan tissue cysts. As tissue cysts were morphologically different from those of Toxoplasma gondii, additional histopathological, immunohistochemical, ultrastructural, and biomolecular investigations were performed, aiming to fully characterize the organism. Histopathology revealed the presence of large Sarcocystis-like tissue cysts, associated with limited inflammatory lesions in all CNS areas studied. IHC was inconclusive, as positive staining with polyclonal antisera did not preclude cross-reaction with other Sarcocystidae coccidia. Applied to each animal, 11 different PCR protocols precluded a neural infection by Sarcocystis neurona, Sarcocystis falcatula, Hammondia hammondi, and Neospora caninum. T. gondii coinfection was confirmed only in dolphin SD2. Sarcocystis sp. sequences, showing the highest homology to species infecting the Bovidae family, were amplified from SD1 myocardium and SD2 skeletal muscle. The present study represents the first report of Sarcocystis-like tissue cysts in the brain of stranded cetaceans along with the first description of Sarcocystis sp. infection in muscle tissue of dolphins from the Mediterranean basin.

Production and use of monoclonal antibodies for identification of Bradyrhizobium japonicum strains

1988 ◽  
Vol 34 (1) ◽  
pp. 88-92 ◽  
Author(s):  
D. Velez ◽  
J. D. Macmillan ◽  
L. Miller
Keyword(s):  
Monoclonal Antibodies ◽  
Bradyrhizobium Japonicum ◽  
Chemical Nature ◽  
Periodate Oxidation ◽  
Enzyme Linked Immunosorbent Assay ◽  
Gram Negative Bacteria ◽  
Bacterial Cells ◽  
Gram Negative ◽  
Polyclonal Antisera ◽  
Somatic Antigens

Thirteen murine hybridomas capable of producing monoclonal antibodies to somatic antigens on Bradyrhizobium japonicum were developed and an indirect enzyme-linked immunosorbent assay was used to test reactivity of the antibodies against 20 strains of B. japonicum. Although polyclonal antisera from mice immunized with strains of B. japonicum reacted with bacterial cells of all 20 strains, individual monoclonals were more specific. Some antibodies reacted with as few as 2 and one with as many as 11 strains. On the basis of reactivity with the set of 13 monoclonal antibodies, the 20 strains of B. japonicum could be divided arbitrarily into five groups. Three of five monoclonal antibodies tested reacted with bacteroids taken directly from soybean nodules. One monoclonal bound to cells of five species of Rhizobium, but none of the 13 reacted with gram-negative bacteria representing six other genera. Treatment of cells with reagents and heat indicated the chemical nature of the antigens to five of the monoclonals. Antigen reactive with one antibody was destroyed by periodate oxidation indicating that it was a polysaccharide. Two antigens were probably proteins as they could be digested by trypsin and denatured by heat. Two others were inactivated by all three treatments suggesting they were glycoproteins.

Novel leukaemia markers

1995 ◽  
Vol 15 (6) ◽  
pp. 463-468
Author(s):  
Kingsley J. Micklem
Keyword(s):  
Monoclonal Antibodies ◽  
T Cell ◽  
Recombinant Protein ◽  
Follicular Lymphoma ◽  
B Lymphocytes ◽  
Synthetic Peptide ◽  
Paraffin Sections ◽  
Hematologic Disorders ◽  
Polyclonal Antisera ◽  
Chromosomal Defects

Using synthetic peptide or recombinant protein as immunising antigens we have produced monoclonal antibodies and polyclonal antisera directed against targets of particular interest in leukaemia diagnosis. In this way we have prepared reagents which recognise all T or all B lymphocytes in routinely fixed paraffin sections which are unique in this respect. We have also produced monoclonal antibodies to molecules potentially involved in specific neoplastic transformations, implicated by virtue of the involvement of their genes in chromosomal defects in these neoplasms. In particular, we have produced antibodies recognising bcl-2, involved in follicular lymphoma, tal-1, involved in T-cell acute leukaemias and HRX involved in a variety of hematologic disorders. The application of these reagents to diagnosis has so far proved useful. In addition their use outside the field of leukaemia diagnosis has proved to be even more important in some cases.

scholarly journals Production of polyclonal antisera using recombinant coat proteins of Grapevine leafroll-associated virus 2 and Grapevine virus B

2008 ◽  
Vol 43 (10) ◽  
pp. 1405-1411 ◽  
Author(s):  
Paula Radaelli ◽  
Thor Vinícius Martins Fajardo ◽  
Osmar Nickel ◽  
Marcelo Eiras ◽  
Gilvan Pio-Ribeiro
Keyword(s):  
Virus Disease ◽  
Disease Diagnosis ◽  
Coat Proteins ◽  
Grapevine Virus ◽  
Western Blots ◽  
E Coli ◽  
Sds Page ◽  
Grapevine Virus B ◽  
Polyclonal Antisera ◽  
Infected Grapevine

The objective of this work was to produce and characterize specific antisera against Brazilian isolates of Grapevine leafroll-associated virus 2 (GLRaV-2) and Grapevine virus B (GVB), developed from expressed coat proteins (CPs) in Escherichia coli, and to test their possible use for the detection of these two viruses in diseased grapevines. The coat protein (CP) genes were RT-PCR-amplified, cloned and sequenced. The CP genes were subsequently subcloned, and the recombinant plasmids were used to transform E. coli cells and express the coat proteins. The recombinant coat proteins were purified, and their identities were confirmed by SDS-PAGE and Western blot and used for rabbit immunizations. Antisera raised against these proteins were able to recognize the corresponding recombinant proteins in Western blots and to detect GLRaV-2 and GVB in infected grapevine tissues, by indirect ELISA, discriminating healthy and infected grapevines with absorbances (A405) of 0.08/1.15 and 0.12/1.30, respectively. Expressing CP genes can yield high amount of viral protein with high antigenicity, and GLRaV-2 and GVB antisera obtained in this study can allow reliable virus disease diagnosis.

scholarly journals Expression of perforin and serine esterases by human gamma/delta T cells.

1991 ◽  
Vol 173 (2) ◽  
pp. 499-502 ◽  
Author(s):  
H Koizumi ◽  
C C Liu ◽  
L M Zheng ◽  
S V Joag ◽  
N K Bayne ◽  
...  
Keyword(s):  
T Cells ◽  
Nk Cells ◽  
Mechanism Of Action ◽  
Lak Cells ◽  
Gamma Delta T Cells ◽  
Serine Esterase ◽  
Gamma Delta ◽  
Western Blots ◽  
Polyclonal Antisera ◽  
Serine Esterases

gamma/delta T cells have recently been described in association with a number of disorders, including autoimmune diseases. gamma/delta T cells are thought to play a cytotoxic role, but their mechanism of action is not known. Several granule mediators of cytotoxicity, including a pore-forming protein (perforin), and a family of serine esterases, have been isolated from cytotoxic T lymphocytes (CTL), lymphokine-activated killer (LAK) cells, and natural killer (NK) cells. We demonstrate here that gamma/delta T cells also express these mediators. Northern blots show that gamma/delta T cells express perforin, serine esterase 1 (SE 1), and SE 2. Three polyclonal antisera - raised against murine perforin, a peptide composed of amino acids 1-34 of human perforin, and human peforin expressed in bacteria - all reacted with a 70-kD protein in gamma/delta T cells on Western blots. Immunostaining with antiperforin antisera shows that primary gamma/delta T cells also contain perforin. Electron microscopy reveals that the granules of gamma/delta T cells resemble those of CTL, LAK, and NK cells. Gamma/delta T cells also resemble LAK cells in possessing inclusion bodies in their nuclei. These results imply that gamma/delta T cells resemble other cytolytic lymphocytes in their mechanism of action.

Polyclonal antisera specific for the proenzyme form of each calpain

1992 ◽  
Vol 1121 (1-2) ◽  
pp. 47-53 ◽  
Author(s):  
Dorothy E. Croall ◽  
Clive A. Slaughter ◽  
Helen S. Wortham ◽  
Colleen M. Skelly ◽  
Lynn DeOgny ◽  
...  
Keyword(s):  
Polyclonal Antisera
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