共查询到20条相似文献,搜索用时 62 毫秒
1.
Eight monoclonal antibodies produced against haemocytes of shrimp (Litopenaeus vannamei) were used to research the antigenic cross-reactivity of crustacean haemocytes. 2C3 cross-reacted with the haemocytes of all the experimental animals, while 1H8 and 2C11 did not cross-react with the experimental animals. The other five monoclonal antibodies cross-reacted with some of the experimental animals. 相似文献
2.
NORMAN E. WILLIAMS JERRY E. HONTS VIRGINIA M. DRESS E. MARLO NELSEN JOSEPH FRANKEL 《The Journal of eukaryotic microbiology》1995,42(4):422-427
ABSTRACT. Twelve monoclonal antibodies were raised that are specific for the membrane skeleton of Tetrahymena . Five were directed against T. pyrifomis and seven were directed against T. thermophila . Some cross-reactivity between species was found. Each monoclonal antibody recognized one of the three major components of epiplasm, i.e. the bands A, B, and C identified in electrophoretic separations of epiplasmic proteins. It was found, using these antibodies, that the epiplasmic proteins A, B, and C have overlapping but independent distributions within the cell. 相似文献
3.
The cucumber long hypocotyl mutant lacks a light-stable PHYB-like phytochrome. 总被引:20,自引:6,他引:14
下载免费PDF全文
![点击此处可从《The Plant cell》网站下载免费的PDF全文](/ch/ext_images/free.gif)
E Lpez-Juez A Nagatani K Tomizawa M Deak R Kern R E Kendrick M Furuya 《The Plant cell》1992,4(3):241-251
A novel cDNA sequence homologous to a phytochrome B (phyB) gene that was isolated in a library from tobacco tissue has been used in an Escherichia coli expression system to raise anti-phytochrome B (anti-PHYB) polypeptide-specific monoclonal antibodies. The specificity of these antibodies has been tested by cross-reactivity against purified pea light-labile type 1 and light-stable type 2 phytochromes, with some antibodies reacting with the type 2 and none with the type 1 phytochromes. One such antibody, monoclonal mAT1, has been employed to analyze the phytochrome molecular species present in a photomorphogenic long hypocotyl (lh) mutant of cucumber. The results indicated that the mutant contains wild-type levels of the light-labile type 1 phytochrome polypeptide (PHYA), which has an apparent molecular mass of approximately 120 kD, but shows less than 1% (detection limit) of a light-stable polypeptide recognized by mAT1 in wild-type seedlings. This protein, not detectable in the lh mutant, has the properties of light-stable type 2 phytochrome, has an apparent molecular mass of 116 to 117 kD, and remains at constant levels under continuous low-fluence-rate red light. Therefore, we conclude that the lh mutant lacks at least one type 2 phytochrome-like polypeptide, most probably a phyB gene product. The correlation between the lack of this protein and the deficiency or absence of physiological responses to a light-stable phytochrome species in this mutant helps to identify the physiological roles played by the products of different subfamilies within the phytochrome gene family. 相似文献
4.
ANGELA HAMPSHIRE C. S. LOPES DIANE McMAHON-PRATT 《The Journal of eukaryotic microbiology》1989,36(4):354-361
Twenty-six monoclonal antibodies were produced against membrane-enriched preparations of Endotrypanum schaudinni or Endotrypanum sp. promastigotes. Fifteen of these monoclonal antibodies (E1-E15) reacted only with the standard strain of E. schaudinni , M6159. Monoclonal antibodies E16-E26 were considered Endotrypanum specific; no cross reactivity was detected with any other genus of the family Trypanosomatidae (Leishmania, Trypanosoma, Leptomonas. Herpetomonas or Crithidia) by dot-blot radioimmune assay. By indirect immunofluorescence assay, the antigens recognized by Endotrypanum specific monoclonal antibodies appear to be associated with the surface of the parasite. Based on Western blot analysis, 4 antigenic molecules ranging in molecular weight from 24 kD to 160 kD were identified by monoclonal antibodies specific for the strain of E. schaudinni , M6159. Monoclonal antibodies specific for the genus Endotrypanum identified an antigen of molecular weight 48 kD as well as a diffuse component migrating with an apparent molecular weight of 64–200 kD. 相似文献
5.
The influence of insect juvenile hormone agonists on metamorphosis and reproduction in estuarine crustaceans 总被引:2,自引:1,他引:1
McKenney CL 《Integrative and comparative biology》2005,45(1):97-105
Comparative developmental and reproductive studies were performedon several species of estuarine crustaceans in response to threejuvenile hormone agonists (pyriproxyfen, methoprene and fenoxycarb).Larval development of the grass shrimp, Palaemonetes pugio,was greater than two orders of magnitude more sensitive to disruptionby methoprene and fenoxycarb than was embryonic development.Developing larvae of the mud crab, Rhithropanopeus harrisii,exhibited reduced metamorphic success at lower concentrationsof methoprene and pyriproxyfen than grass shrimp larvae. Theseresponses suggest that the more rigidly controlled metamorphicprocess in crabs is more sensitive to compounds acting as endocrinedisruptors than is the more flexible metamorphic pattern inshrimp. The final crab larval stage, the megalopa, was moresensitive to methoprene and fenoxycarb exposure than earlierzoeal stages. Mud crab larvae exposed to fenoxycarb had reducedbiomass and lipid content, particularly triglycerides and sterols.Concentrations of fenoxycarb which reduced the reproductivecapacity in single life-cycle exposures of the estuarine mysid,Americamysis bahia, were similar to those concentrations whichinhibited metamorphosis in grass shrimp. Juvenile mysids releasedby exposed adults and reared through maturation without furtherexposure produced fewer young and had altered sex ratios (lowerpercentages of males) at lower parental-exposure concentrationsthan directly affected parental reproduction. These transgenerationalresponses may well be a product of irreversible effects duringdevelopmental exposures which become apparent following maturationand initiation of reproduction. These findings support usinga functional approach as an appropriate screening procedureto evaluate potential environmental endocrine-disrupting chemicalsin aquatic environments. 相似文献
6.
Twenty-six monoclonal antibodies were produced against membrane-enriched preparations of Endotrypanum schaudinni or Endotrypanum sp. promastigotes. Fifteen of these monoclonal antibodies (E1-E15) reacted only with the standard strain of E. schaudinni, M6159. Monoclonal antibodies E16-E26 were considered Endotrypanum specific; no cross reactivity was detected with any other genus of the family Trypanosomatidae (Leishmania, Trypanosoma, Leptomonas, Herpetomonas or Crithidia) by dot-blot radioimmune assay. By indirect immunofluorescence assay, the antigens recognized by Endotrypanum specific monoclonal antibodies appear to be associated with the surface of the parasite. Based on Western blot analysis, 4 antigenic molecules ranging in molecular weight from 24 kD to 160 kD were identified by monoclonal antibodies specific for the strain of E. schaudinni, M6159. Monoclonal antibodies specific for the genus Endotrypanum identified an antigen of molecular weight 48 kD as well as a diffuse component migrating with an apparent molecular weight of 64-200 kD. 相似文献
7.
8.
Calmodulin-dependent protein phosphatase (CaM-PPase) was isolated from bovine parotid gland by sequential application of DEAE-52, Affi-gel blue and calmodulin-affinity chromatography followed by gel filtration and high performance liquid chromatography. The enzyme was activated in the simultaneous presence of Ni2+ or Mn2+ and Ca2+ plus calmodulin. Ca2+/calmodulin-dependent activation of CaM-PPase was antagonized by inhibitors of calmodulin action, such as W-7 and trifluoperazine. Tryptophan fluorescence was quenched in the presence of Ni2+. CaM-PPase was a heterodimer. The molecular weights of large subunits which bound calmodulin (CaM) were 68 kD and 58 kD - the 68 kD subunit was predominant. Polyclonal antibodies against bovine calcineurin cross-reacted with both types of larger subunits. Using polyclonal antibodies against bovine calcineurin or the monoclonal antibody against subunit B of bovine calcineurin, the smaller molecular weight subunit (19 kD) was found to be immunologically identical to subunit B of bovine calcineurin. In bovine parotid gland, CaM-PPase was found both in acinar and duct cells. 相似文献
9.
不同水生脊椎动物的血清与黏液蛋白图谱分析 总被引:1,自引:0,他引:1
因脊椎动物的血清与黏液中含有大量蛋白质及其他生物活性物质, 而具有多种生理、生化与抗病防御功能。为了解水生脊椎动物血清与黏液中所含主要蛋白成分及其异同, 我们分别从中华鲟(Chinese sturgeonAclpenser sinensis grdy)、鲫鱼(Crucian carp Carassius auratus)、草鱼(Grass carp Ctenopharyngodonidellus)、赤点石斑鱼(Red-spotted grouper Epinephelus akaara)、青石斑鱼(Banded grouper Epinephelusawoara)、狗鱼(Pikes Esox reicherti)、江豚(Finless porpoise Neophocaena phocaenoides)、黄颡鱼(Yellow catfishPelteobagrus fulvidraco)、鳜鱼(Mandarin fish Siniperca chuatsi) 这9 种水生脊椎动物中, 采集血清和皮肤黏液样本。利用考马斯亮蓝G-250 法, 测定了不同水生脊椎动物的血清和黏液样品中总蛋白的含量; 经SDS-PAGE 蛋白电泳, 分别获得不同水生脊椎动物血清及黏液蛋白图谱; 比较和分析了同种动物血清和黏液、不同动物之间血清与血清或黏液与黏液之间蛋白图谱的差异。结果表明:这些动物的血清蛋白组分有显著相似性, 在分子量45—120 kD 之间, 蛋白条带大量集中分布; 除青石斑鱼外, 其他8 种水生脊椎动物都有分子量约为 28 kD 和 14 kD 相同或相近的两条蛋白带。草鱼、鲫鱼、鳜鱼、狗鱼、黄颡鱼和江豚的黏液蛋白, 除了都含有分子量大小约为45 kD 的蛋白条带外, 其他无显著相似性。对同种水生脊椎动物的血清与黏液蛋白比较, 虽然黄颡鱼的血清与黏液中分子量相近的蛋白带最多, 也仅约为38%, 可见其蛋白带的种类和含量都存在显著差异。
相似文献
10.
Dolgova NV Kamanina IuV Akimova OA Orlov SN Rubtsov AM Lopina OD 《Biochemistry. Biokhimii?a》2007,72(8):863-871
Immunoprecipitation of Na,K-ATPase from kidney homogenate by antibodies against alpha1-subunit results in the precipitation of several proteins together with the Na,K-ATPase. A protein with molecular mass of about 67 kD interacting with antibodies against melittin (melittin-like protein, MLP) was found in the precipitate when immunoprecipitation was done in the presence of ouabain. If immunoprecipitation was done using antibodies against melittin, MLP and Na,K-ATPase alpha1-subunit were detected in the precipitate, and the amount of alpha1-subunit in the precipitate was increased after the addition of ouabain to the immunoprecipitation medium. MLP was purified from mouse kidney homogenate using immunoaffinity chromatography with antibodies against melittin. The addition of MLP to purified FITC-labeled Na,K-ATPase decreases fluorescence in medium with K+ and increases it in medium with Na+. The enhancement of fluorescence depends upon the MLP concentration. The N-terminal sequence of MLP determined by the Edman method is the following: HPPKRVRSRLNG. No proteins with such N-terminal sequence were found in the protein sequence databases. However, we revealed five amino acid sequences that contain this peptide in the middle part of the chain at distance 553 amino acids from the C-terminus (that corresponds to protein with molecular mass of about 67 kD). Analysis of amino acid sequence located between C-terminus and HPPKRVRSRLNG in all found sequences has shown that they were highly conservative and include WD40 repeats. It is suggested that the 67-kD MLP either belongs to the found protein family or was a product of proteolysis of one of them. 相似文献
11.
12.
The present study disclosed the cross-reactivity between Bermuda grass pollen (BGP) and other grass pollens using monoclonal antibodies (MAbs) and polyclonal antiserum. MAb 9–13, directed against a group of minor allergens of BGP (Cyn d Bd68K, 48K, 38K) was found to cross-react with extracts of ten other grass pollens. Immunoblotting assays illustrated that MAb 9–13 cross-reacted with multiple components of most of these pollens, and the major cross-reactive components had molecular weights of 29–36 kD. The cross-reactivity between BGP andLol pI, the group I allergen of rye grass pollen, was further evaluated;Lol pI was recognized by MAb 9–13, but not by our MAbs/polyclonal antiserum againstCyn dI, the major allergen of BGP. These results suggest that the epitope recognized by MAb 9–13 is a common (C) epitope shared byLol pI andCyn d Bd68K, 48K, 38K, andCyn dI does not share significant antigenicity withLol pI. In a modified radio-allergosorbent test, IgE antibodies in the serum of BGP-allergic patients reacted mildly with C-epitope-bearing components of both BGP and rye grass pollens, and this binding could be blocked specifically by MAb 9–13. This suggests that in addition to an antigenic cross-reaction, the C epitope can also lead to an allergenic cross-reaction. 相似文献
13.
Phylogenetic cross-reactivities of monoclonal antibodies produced against rat neurophysin 总被引:2,自引:0,他引:2
Yakov Ben-Barak James T. Russell Mark Whitnall Keiko Ozato Harold Gainer 《Cellular and molecular neurobiology》1984,4(4):339-349
Previously described mouse monoclonal antibodies against rat neurophysins [Ben-Barak, Y., et al. (1985); Whitnall, M. H., et al. (1985)] were studied here for their cross-reactivities to neurophysins (NPs) from other vertebrate species. Posterior pituitary extracts from various mammals (rat, mouse, cow, human) and lower vertebrates (frog, ratfish) were studied. The monoclonal antibodies displayed several distinct patterns of cross-reactivity to the various species, indicating that the epitopes which they recognized were different. PS 67 bound strongly to rat pituitary extract in solid-phase radioimmunoassay (RIA) but showed no cross-reactivity with extracts from any of the other species tested, including the mouse. PS 36 cross-reacted with mouse and frog extracts but showed almost no cross-reactivity with cow and none to ratfish extracts. PS 41 cross-reacted with mouse, cow, and frog extracts. PS 45 was the most cross-reactive antibody and recognized an antigen in extracts from mouse, cow, frog, and ratfish pituitaries. Electrophoresis of proteins extracted from posterior pituitaries, followed by immunoblot staining with either PS 36 or PS 45, demonstrated that the NP-like molecules within each species are heterogeneous, i.e., more than two bands stained in each species. The frog NP stained by PS 45 was about twice the molecular weight of the mammalian NPs. The possible valve of the PS 45 antibody for future molecular cloning experiments on the arginine vasotocin precursor in lower vertebrates is discussed. 相似文献
14.
Zhijian Tu David N. Byrne Henry H. Hagedorn 《Archives of insect biochemistry and physiology》1997,34(2):223-237
SDS-PAGE of the sweet potato whitefly (Bemisia tabaci) egg extract showed one major band (approximately 190 kDa) and two minor bands (approximately 75 kDa and 67 kDa). A distinct 190 kDa band was also present in male extract. On SDS gels the vitellin band of the greenhouse whitefly (Trialeurodes vaporarium) was larger, about 220 kDa. The native molecular mass of sweet potato whitefly vitellin was estimated to be 375 kDa using 4–20% native pore-limiting gel electrophoresis. Its isoelectric point was estimated to be 7.3 using isoelectric focusing. Two-dimensional gel electrophoresis and densitometry were used to estimate vitellin subunit composition; the data suggest that the sweet potato whitefly vitellin is likely to be a 380 kDa native molecule formed by two 190 kDa subunits. The two minor bands (75 kDa and 67 kDa) may be breakdown products of the native vitellin. This conclusion was supported by a Western blot of an SDS-PAGE gel of partially degraded female and egg extracts, which showed that polyclonal antiserum raised against the 190 kDa polypeptide recognized the 75 kDa and 67 kDa bands. Seven hybridoma cell lines secreting monoclonal antibodies against the 190 kDa band were screened, and one of them (S1A2G9H2) was mass produced. The antibody recognized the 190 kDa band in a Western blot. All the screened monoclonal antibodies were female and egg-specific by ELISA and/or Western blot, suggesting that the 190 kDa band in male extract was not a vitellin. A sensitive ELISA was established that could detect as little as 1/40 of an egg equivalent of vitellin using the monoclonal antibody from S1A2G9H2. Profiles of female sweet potato whitefly reproductive activities (egg laying, amount of vitellin in the female, and total vitellin produced by a female) within 2 days after eclosion were determined. Arch. Insect Biochem. Physiol. 34:223–237, 1997. © 1997 Wiley-Liss, Inc. 相似文献
15.
Characterization of components of Z-bands in the fibrillar flight muscle of Drosophila melanogaster 总被引:12,自引:4,他引:8
下载免费PDF全文
![点击此处可从《The Journal of cell biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
J D Saide S Chin-Bow J Hogan-Sheldon L Busquets-Turner J O Vigoreaux K Valgeirsdottir M L Pardue 《The Journal of cell biology》1989,109(5):2157-2167
Twelve monoclonal antibodies have been raised against proteins in preparations of Z-disks isolated from Drosophila melanogaster flight muscle. The monoclonal antibodies that recognized Z-band components were identified by immunofluorescence microscopy of flight muscle myofibrils. These antibodies have identified three Z-disk antigens on immunoblots of myofibrillar proteins. Monoclonal antibodies alpha:1-4 recognize a 90-100-kD protein which we identify as alpha-actinin on the basis of cross-reactivity with antibodies raised against honeybee and vertebrate alpha-actinins. Monoclonal antibodies P:1-4 bind to the high molecular mass protein, projectin, a component of connecting filaments that link the ends of thick filaments to the Z-band in insect asynchronous flight muscles. The anti-projectin antibodies also stain synchronous muscle, but, surprisingly, the epitopes here are within the A-bands, not between the A- and Z-bands, as in flight muscle. Monoclonal antibodies Z(210):1-4 recognize a 210-kD protein that has not been previously shown to be a Z-band structural component. A fourth antigen, resolved as a doublet (approximately 400/600 kD) on immunoblots of Drosophila fibrillar proteins, is detected by a cross reacting antibody, Z(400):2, raised against a protein in isolated honeybee Z-disks. On Lowicryl sections of asynchronous flight muscle, indirect immunogold staining has localized alpha-actinin and the 210-kD protein throughout the matrix of the Z-band, projectin between the Z- and A-bands, and the 400/600-kD components at the I-band/Z-band junction. Drosophila alpha-actinin, projectin, and the 400/600-kD components share some antigenic determinants with corresponding honeybee proteins, but no honeybee protein interacts with any of the Z(210) antibodies. 相似文献
16.
N C Engleberg E Pearlman D Dixon B I Eisenstein 《Journal of immunology (Baltimore, Md. : 1950)》1986,136(4):1415-1417
We studied the antigenic cross-reactivity of surface proteins among various strains of Legionella pneumophila and other Legionella species by using a novel method of antibody purification. Anti-bacterial antibodies in hyperimmune sera were adsorbed to and eluted from the surface of recombinant E. coli cells that express individual L. pneumophila antigens on their surface. These affinity-purified antibodies were then used to probe protein immunoblots prepared from the test strains to detect cross-reactive domains. We found that antigenic proteins are generally conserved in all L. pneumophila serogroups. Although some of these antigenic domains are shared with members of other Legionella species, they are associated with proteins of different molecular mass. Our approach to the study of antigenic cross-reactivity has potential advantages over similar studies that use either monoclonal antibodies or monospecific antibodies prepared by immunization with purified antigens. 相似文献
17.
锯缘青蟹主要过敏原的纯化与鉴定 总被引:2,自引:0,他引:2
以锯缘青蟹为研究对象,从免疫鉴定、分离纯化、抗体制备和免疫学分析等方面对其主要过敏原进行研究。首先利用过敏者血清的免疫印迹法,确定锯缘青蟹的主要过敏原为分子量约38kD的蛋白。然后通过制备丙酮粉、等电点沉淀、硫酸铵沉淀及加热处理对分子量为38kD的主要过敏蛋白进行了高度纯化。该蛋白的pI约为4.5,与虾的原肌球蛋白Pena1性质相近,证实了锯缘青蟹的主要过敏原为原肌球蛋白。通过免疫新西兰大白兔,制备了原肌球蛋白的抗血清,采用Protein A Sepharose亲和层析柱对动物抗体进行了纯化。该抗血清效价高,经4×105倍稀释后仍能与抗原进行反应。该抗体与甲壳类动物及软体动物的原肌球蛋白具有较强的免疫交叉反应,可用于食品过敏原检测。
相似文献
18.
K. P. Liu P.-Y. Yu S. H. Hsiung A. L. Kirchgessner† M. D. Gershon† H. Tamir† 《Journal of neurochemistry》1990,55(3):1013-1021
Serotonin binding protein (SBP) is a constituent of the synaptic vesicles of serotonergic neurons. Two types of SBP, with molecular masses of 45 kDa and 56 kDa, have been purified. To determine whether there are shared epitopes between the two forms of SBP, we raised and tested for cross-reactivity monoclonal antibodies (MAbs) against each form of SBP. We obtained 12 MAbs, all of which recognize both forms of SBP. Hybridoma clones were produced by fusing P3 X 63Ag8.653 mouse myeloma cells with spleen cells from a mouse that had been immunized with 45-kDa or 56-kDa SBP. Culture supernatants were screened for the presence of anti-SBP antibodies. MAb isotypes were determined by immunodiffusion, using immunoglobulin type-specific antisera. Each antibody to SBP consisted of only a single subclass of immunoglobulin (IgM). We obtained 12 MAbs, each of which interacted with both forms of SBP, as judged by enzyme-linked immunosorbent assay and immunoblot analysis. Ascites fluid to one clone (44-10) was obtained and affinity-purified. In the presence of goat anti-mouse IgM, the partially purified 44-10 antibodies quantitatively immunoprecipitated SBP from crude brain extracts. Immunoblotting revealed two major bands corresponding to 45 kDa and 56 kDa and a minor band corresponding to 68 kDa. MAb 44-10 blocked the binding of [3H]serotonin ([3H]5-HT) to 45-kDa and 56-kDa SBP in a concentration-dependent manner. The 68-kDa protein was found to bind [3H]5-HT. Sites reacting with MAB 44-10 were located immunocytochemically in sections of rat brain.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
19.
L Monjour A Berneman I Vouldoukis M Domurado M C Guillemin C Chopin C Alfred A Roseto 《Comptes rendus de l'Académie des sciences. Série III, Sciences de la vie》1985,300(9):395-398
Mice monoclonal antibodies (IgG) have been raised against Leishmania infantum promastigotes by fusing SP 2/0 myeloma cells and immunized mice splenic cells. The monoclonal antibodies have been detected by indirect immunofluorescence. In vivo tests showed that some of them could inhibit the life cycle of several Leishmania species from the Old and the New World. Studies of these protective monoclonals by the western blot technique showed the presence of three constant antigens (40 kD, 70 kD and 113 kD) amongst the Leishmania species studied. 相似文献
20.
Kumar B Deepika A Makesh M Purushothaman CS Rajendran KV 《Journal of invertebrate pathology》2012,111(1):86-89
Monoclonal antibodies (MAbs) to hemocytes of mud crab, Scylla serrata, were produced by immunizing mice with formalin-fixed hemocytes. Of the six MAbs produced, two (MAb 1D11 and MAb 1A2) reacted specifically with hemocyte proteins in western blot. MAb 1A2 showed strong immunofluorescent reaction with granular hemocytes and a weak reaction with semigranular cells. However, hyaline cells did not show any reaction. The MAbs also showed strong cross-reactivity with the hemocytes of different crab species but not with other crustaceans. The MAbs produced can be used as a marker for granular hemocytes of crabs. 相似文献