Immunohistochemical Staining of Cyclooxygenases with Monoclonal Antibodies

Author(s):  
Ghassan M. Saed
1988 ◽  
Vol 8 (5) ◽  
pp. 2251-2256
Author(s):  
P De Togni ◽  
H Niman ◽  
V Raymond ◽  
P Sawchenko ◽  
I M Verma

We have generated monoclonal antibodies by using a synthetic peptide corresponding to amino acid positions 4 to 17 of the human fos protein. The antibodies detected both v- and c-fos proteins by immunoprecipitation, immunoblotting, and indirect immunofluorescence. The monoclonal antibodies not only identified the fos protein complex with the cellular 39-kilodalton protein, but also recognized the modified forms of the mouse, rat, and human fos proteins. In day-17 rat embryos, nuclear-staining fos protein could be identified in the cartilage by immunohistochemical staining.


2009 ◽  
Vol 54 (No. 6) ◽  
pp. 270-276
Author(s):  
M. Simon ◽  
J. Antalíková ◽  
Ľ. Horovská ◽  
J. Jankovičová ◽  
K. Fábryová ◽  
...  

Studies that involved testing monoclonal antibodies (mAbs) for cross-species reactivity proved to be efficient for the identification of previously unrecognized antigens in a number of different species. Twenty-six mAbs specific to different bovine CD (cluster defined) antigens (CD9, CD18, CD45R, CD41/61, CD62L, MHC class I and bovine IgG light chain molecule) were assayed for reactivity with rabbit peripheral blood leukocytes. Four of the mAbs recognizing CD9 and CD41/61 were reactive with rabbit platelets or granulocytes. These were investigated further by immunoblotting and immunohistochemical staining. The study identified CD9 and CD41/61 molecules on rabbit cells by mAbs IVA-50 and IVA-38. It showed that IVA-50 is a new valuable CD9 reagent for rabbit immunology which could be used for immunofluorescence staining or ELISA assay, immunohistological and molecular studies of rabbit CD9 antigen. IVA-38 recognizes the CD41/61 on rabbit platelets in indirect immunofluorescence and ELISA assay.


1988 ◽  
Vol 8 (5) ◽  
pp. 2251-2256 ◽  
Author(s):  
P De Togni ◽  
H Niman ◽  
V Raymond ◽  
P Sawchenko ◽  
I M Verma

We have generated monoclonal antibodies by using a synthetic peptide corresponding to amino acid positions 4 to 17 of the human fos protein. The antibodies detected both v- and c-fos proteins by immunoprecipitation, immunoblotting, and indirect immunofluorescence. The monoclonal antibodies not only identified the fos protein complex with the cellular 39-kilodalton protein, but also recognized the modified forms of the mouse, rat, and human fos proteins. In day-17 rat embryos, nuclear-staining fos protein could be identified in the cartilage by immunohistochemical staining.


Author(s):  
Pengfei Wang ◽  
Chunguo Liu ◽  
Shida Wang ◽  
Lili Wen ◽  
Zhibin Shi ◽  
...  

AbstractAfrican swine fever (ASF) is an acute hemorrhagic disease of domestic pigs. The causative agent of ASF, ASF virus (ASFV), is a double-stranded DNA virus, the sole member in the family Asfarviridae. The non-structural protein pB602L of ASFV is a molecular chaperone of the major capsid protein p72 and plays a key role in icosahedral capsid assembly. This protein is antigenic and is a target for developing diagnostic tools for ASF. To generate monoclonal antibodies (mAbs) against pB602L, a prokaryotically expressed recombinant pB602L protein was produced, purified, and used as an antigen to immunize mice. A total of eight mouse mAbs were obtained, and their binding epitopes were screened by Western blot using an overlapping set of polypeptides from pB602L. Three linear epitopes were identified and designated epitope 1 (366ANRERYNY373), epitope 2 (415GPDAPGLSI423), and epitope 3 (498EMLNVPDD505). Based on the epitope recognized, the eight mAbs were placed into three groups: group 1 (B2A1, B2F1, and B2D10), group 2 (B2H10, B2B2, B2D8, and B2A3), and group 3 (B2E12). The mAbs B2A1, B2H10, and B2E12, each representing one of the groups, were used to detect pB602L in ASFV-infected porcine alveolar macrophages (PAMs) and pig tissues, using an indirect fluorescence assay (IFA) and immunohistochemical staining, respectively. The results showed that pB602L was detectable with all three mAbs in immunohistochemical staining, but only B2H10 was suitable for detecting the proteins in ASFV-infected PAMs by IFA. In summary, we developed eight anti-pB602L mouse mAbs recognizing three linear epitopes in the protein, which can be used as reagents for basic and applied research on ASFV.


1995 ◽  
Vol 10 (3) ◽  
pp. 129-135 ◽  
Author(s):  
L.P. Howell ◽  
S.J. Denardo ◽  
N.B. Levy ◽  
J. Lund ◽  
G.L. Denardo

Five monoclonal antibodies (MoAbs) (L6, 170H.82, 155, BrE-3 and BR96), most of which have been previously shown to target breast cancer and not normal tissues by immunoscintigraphic imaging, were evaluated for their frequency and pattern of immunohistochemical staining in 67 to 116 metastatic lesions from patients with ductal carcinoma of the breast. Immunoperoxidase staining in 75% or more of the cells occurred in 56/116 (48%) for L6, 44189 (49%) for Br, -96, 58/102 (57%) for 155, 62/99 (84%) for 170H.82, and 65.67 (97%) for BrE-3. With the first three MoAbs, an additional 6-10% of the tumors showed staining in 50-75% of tumor cells. These results illustrate that most patients with metastatic ductal carcinoma have cancer tissue in which a high percent of cells will react to several of these selected MoAbs that target different epitopes. The high expression of the MoAb targets throughout the tumor tissue makes these antibodies potential candidates to carry immunologically directed radioimmunotherapy and is an aid in selecting patients for treatment..


2021 ◽  
Author(s):  
Mi-Jin Park ◽  
Jongjin Park ◽  
Slki Park ◽  
Sunghwa Choe

AbstractThe biotechnological applications of the programmable DNA-cleaving enzymes known as clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) continue to expand from agricultural trait development to therapeutic gene therapies. The Cas9 CRISPR system isolated from Streptococcus pyogenes (SpyCas9) is classified as class II, which is popular due to high efficiency and robustness. Antibodies that specifically detect SpyCas9 could expand the applicability of this enzyme. Here, we report on the development of monoclonal antibodies against SpyCas9. Four hybridoma cells were selected for their expression of anti-SpyCas9. Hybridoma supernatant contained reactive and highly specific antibodies to SpyCas9. Anti-SpyCas9 antibody was purified and was non-selective against six other bacterial Cas9 proteins. SpyCas9 protein could be detected in HEK293T cells in decreasing amounts over a 48 hour period, indicating the antibody could be used to detect residual levels of SpyCas9 remaining following cell treatment with a CRISPR/SpyCas9 system. The anti-SpyCas9 antibody may help researchers facilitate Cas9 for further study for the use of techniques such as enzyme-linked immunosorbent assays (ELISAs), western blot, immunoprecipitation, and immunohistochemical staining.


2010 ◽  
Vol 56 (2) ◽  
pp. 36-41
Author(s):  
Iu M Kolesnik ◽  
A M Kamyshnyĭ ◽  
A V Abramov ◽  
O V Gancheva ◽  
V A Liubomirskaia

The objective of the present work was to study the morphofunctional state of insulin-expressing, antigen-presenting, Treg cells, epithelioreticulocytes, thymic cells expressing c-Fos and Bcl-2+ in the progeny of rats with experimental gestational diabetes (EGD). The methods employed included immunohistochemical staining with direct and indirect immunofluorescence using monoclonal antibodies against insulin, MHC-II antigen, CD25 antigen, cytokeratins, rat Bcl-2 and c-Fos proteins. The off-springs of rats with experimental gestational diabetes were shown to have multiple disturbances of immunoregulation in the thymus, such as abnormalities of Treg cell differentiation, impaired expression of early response proteins (c-Fos), reduced number of epithelioreticulocytes, antigen-presenting and insulin-immunopositive cells, hyperexpression of anti-apoptotic Bcl-2 protein. These changes interfere with the normal apoptotic process, selection of thymocytes, and development of central tolerance to pancreatic antigens.


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