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MOUSE ANTI HUMAN FIBRINOPEPTIDE A

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[#ABS11685] MOUSE ANTI HUMAN FIBRINOPEPTIDE A

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(1) Coagulant thrombin-like enzyme (barnettobin) from Bothrops barnetti venom: molecular sequence analysis of its cDNA and biochemical properties.[TOP]

Pubmed ID :23578498
Publication Date : //
The thrombin-like enzyme from Bothrops barnetti named barnettobin was purified. We report some biochemical features of barnettobin including the complete amino acid sequence that was deduced from the cDNA. Snake venom serine proteases affect several steps of human hemostasis ranging from the blood coagulation cascade to platelet function. Barnettobin is a monomeric glycoprotein of 52 kDa as shown by reducing SDS-PAGE, and contains approx. 52% carbohydrate by mass which could be removed by N-glycosidase. The complete amino acid sequence was deduced from the cDNA sequence. Its sequence contains a single chain of 233 amino acid including three N-glycosylation sites. The sequence exhibits significant homology with those of mammalian serine proteases e.g. thrombin and with homologous TLEs. Its specific coagulant activity was 251.7 NIH thrombin units/mg, releasing fibrinopeptide A from human fibrinogen and showed defibrinogenating effect in mouse. Both coagulant and amidolytic activities were inhibited by PMSF. N-deglycosylation impaired its temperature and pH stability. Its cDNA sequence with 750 bp encodes a protein of 233 residues. Indications that carbohydrate moieties may play a role in the interaction with substrates are presented. Barnettobin is a new defibrinogenating agent which may provide an opportunity for the development of new types of anti-thrombotic drugs.

Authors : Vivas-Ruiz Dan E, Sandoval Gustavo A, Mendoza Julio, Inga Rosalina R, Gontijo Silea, Richardson Michael, Eble Johannes A, Yarleque Armando, Sanchez Eladio F,



(2) Proliferating and helper T lymphocytes display distinct fine specificities in response to human fibrinopeptide B.[TOP]

Pubmed ID :6189893
Publication Date : //
The fine specificity of T cell responses involved in the generation of help for antibody production and proliferation was examined by using the 14 amino acid peptide human fibrinopeptide B (hFPB, B beta 1-14) and its synthetic peptide homologues B beta 1-14(Lys14), B beta 1-13, and B beta 3-14. Peritoneal exudate or lymph node T cells from C57BL/10 and B10.BR mice immunized with hFPB or its synthetic homologues were used to measure in vitro proliferative responses. T cells from hFPB-immunized B10.BR mice showed specific proliferation to hFPB, but were unresponsive to B beta 1-14(Lys14), B beta 1-13, and B beta 3-14. B10.BR mice immunized with B beta 1-14(Lys14), B beta 1-13, or B beta 3-14 were unresponsive to all peptides tested. T cells from C57BL/10 mice showed no specific proliferation after immunization and challenge with any of the peptide antigens. In contrast to the patterns of T cell proliferation, immunization of both B10.BR and C57BL/10 mice with hFPB, B beta 1-14(Lys14), B beta 1-13, or B beta 3-14 primed for significant helper T cell activity, as assessed by the augmentation of a primary in vitro B cell IgM anti-FITC plaque-forming cell response after culture with B beta 1-1(Lys14)-FITC. Significant peptide-specific helper activity was observed when the FITC moiety was conjugated to the carboxyl terminal lysine (B beta 1-14(Lys14)-FITC) as well as FITC substitution at the amino terminus (FITC-B beta 3-13 or FITC-B beta 3-14). These results suggest that the fine specificity of T cell responses to peptide antigens are different for helper and proliferating T cells and that responsiveness by one T cell subpopulation does not predict the response pattern of other functional subpopulations.

Authors : Peterson L B, Wilner G D, Thomas D W,



(3) Functional differentiation in the genetic control of murine T lymphocyte responses to human fibrinopeptide B.[TOP]

Pubmed ID :6217245
Publication Date : //
The ability to generate proliferative and helper T lymphocyte responses in mice was compared by using the 14 amino acid peptide, human fibrinopeptide B (hFPB). Lymph node or peritoneal exudate T cells from mice immunized with hFPB were assessed for in vitro proliferation to soluble hFPB as determined by the uptake of 3H-thymidine. The T cell proliferative response to hFPB was found to be under MHC-linked Ir gene control; mice possessing the H-2a,k haplotypes were responders, whereas H-2b,d,q,s mice were nonresponders. The influence of non-H-2 genes on these responses was not investigated, so exclusive regulation by H-2 is provisional. The absence of a detectable lymph node and peritoneal exudate T cell proliferative response persisted in H-2b,d,q,s mice after immunization and boosting with several doses of hFPB. In addition, the capacity to produce a T cell proliferative response was inherited in an autosomal dominant manner and gene(s) controlling responsiveness to hFPB mapped to the I-A subregion of the H-2 complex. To measure peptide-specific helper T cell activity, an in vitro microculture assay in which hFPB-primed lymph node T cells and normal spleen B cells and macrophages were used was developed measuring anti-fluorescein isothiocyanate (FITC) IgM and IgG plaque-forming cell (PFC) responses after culture with FITC-conjugated peptide. Immunization of B10.BR, C57BL/10, B10.D2, and B6AF mice with hFPB primed for significant helper T cell activity as assessed by the ability to augment a primary in vitro IgM response to FITC. The normal B cell IgM responses were completely dependent on hFPB-primed T cells and required that hapten (FITC) and carrier (peptide) be linked. In addition, immunization with FITC-conjugated peptide elicited positive in vivo PFC responses to FITC in B10.BR and C57BL/10 mice, indicating similar genetic control of helper activity in both the intact animals and the in vitro microcultures. Thus, B10.BR mice show both T help and T proliferative responses to hFPB, whereas C57BL/10 mice show only T help and no T proliferative responses. In contrast to B10.BR mice, C3H and CBA mice immunized with hFPB were completely unresponsive when assayed for helper T cell activity in vitro despite their ability to generate positive lymph node T cell proliferative responses. These results indicate responsiveness to hFPB by T helper and proliferating cells is different and is under separate genetic control.

Authors : Peterson L B, Wilner G D, Thomas D W,