Preparation of rabbit polyclonal antibody against porcine gasdermin D protein and determination of the expression of gasdermin D in cultured cells and porcine tissues

2021 ◽  
pp. 105945
Author(s):  
Jiameng Song ◽  
Kexin Wang ◽  
Bo Ma ◽  
Junwei Wang ◽  
Wenlong Zhang
Keyword(s):  
Polyclonal Antibody ◽  
Cultured Cells ◽  
Rabbit Polyclonal Antibody ◽  
Porcine Tissues

scholarly journals An SPRi Biosensor for Determination of the Ovarian Cancer Marker HE4 in Human Plasma

Sensors ◽  
2021 ◽  
Vol 21 (10) ◽  
pp. 3567
Author(s):  
Beata Szymanska ◽  
Zenon Lukaszewski ◽  
Beata Zelazowska-Rutkowska ◽  
Kinga Hermanowicz-Szamatowicz ◽  
Ewa Gorodkiewicz
Keyword(s):  
Ovarian Cancer ◽  
Cancer Patients ◽  
Tumor Resection ◽  
Analytical Signal ◽  
Cancer Marker ◽  
Human Epididymis Protein 4 ◽  
Pearson Coefficient ◽  
Rabbit Polyclonal Antibody ◽  
Good Agreement

Human epididymis protein 4 (HE4) is an ovarian cancer marker. Various cut-off values of the marker in blood are recommended, depending on the method used for its determination. An alternative biosensor for HE4 determination in blood plasma has been developed. It consists of rabbit polyclonal antibody against HE4, covalently attached to a gold chip via cysteamine linker. The biosensor is used with the non-fluidic array SPRi technique. The linear range of the analytical signal response was found to be 2–120 pM, and the biosensor can be used for the determination of the HE4 marker in the plasma of both healthy subjects and ovarian cancer patients after suitable dilution with a PBS buffer. Precision (6–10%) and recovery (101.8–103.5%) were found to be acceptable, and the LOD was equal to 2 pM. The biosensor was validated by the parallel determination of a series of plasma samples from ovarian cancer patients using the Elecsys HE4 test and the developed biosensor, with a good agreement of the results (a Pearson coefficient of 0.989). An example of the diagnostic application of the developed biosensor is given—the influence of ovarian tumor resection on the level of HE4 in blood serum.

Determination of the Viability of Toxoplasma gondii in Cured Ham Using Bioassay: Influence of Technological Processing and Food Safety Implications

2010 ◽  
Vol 73 (12) ◽  
pp. 2239-2243 ◽  
Author(s):  
SUSANA BAYARRI ◽  
MARÍA J. GRACIA ◽  
REGINA LÁZARO ◽  
CONSUELO PÉREZ-ARQUILLUÉ ◽  
MONTSERRAT BARBERÁN ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Horizontal Transmission ◽  
Meat Product ◽  
Immunofluorescence Assay ◽  
Nitrite Concentration ◽  
Bioassay Technique ◽  
Technological Processing ◽  
Undercooked Meat ◽  
Porcine Tissues

Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii and distributed worldwide. Ingestion of viable cysts from infected raw or undercooked meat is an important route of horizontal transmission of the parasite to humans. Little information is available concerning the effect of commercial curing on cysts of T. gondii. This study is the first in which the influence of processing of cured ham on the viability of T. gondii has been evaluated, using bioassay to assess the risk of infection from eating this meat product. Naturally infected pigs were selected for the study, and a mouse concentration bioassay technique was used to demonstrate viable bradyzoites of T. gondii in porcine tissues and hams. No viable parasites were found in the final product (14 months of curing) based on results of the indirect immunofluorescence assay and histological and PCR analyses. Our results indicate that the consumption of hams cured as described here poses an insignificant risk of acquiring toxoplasmosis. However, additional studies are required to evaluate the safety of ham products cured under different conditions of curing time, salt, and nitrite concentration.

An immunological determination of somatostatin in pharmaceutical by sandwich ELISA based on IgY and polyclonal antibody

2019 ◽  
Vol 145 ◽  
pp. 532-538
Author(s):  
Yingshan Chen ◽  
Yuxin Liang ◽  
Rui Lv ◽  
Nana Xia ◽  
Tianjian Xue ◽  
...  
Keyword(s):  
Polyclonal Antibody ◽  
Sandwich Elisa

scholarly journals The correlation between IL-10 expression and histopathological type in nasopharynx carcinoma patients

2020 ◽  
Vol 50 (1) ◽  
pp. 52
Author(s):  
Hamita Hamita ◽  
Muhtarum Yusuf ◽  
Manshur Shidiq Wiyadi
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Polyclonal Antibody ◽  
Therapy Response ◽  
P Value ◽  
Fisher Exact Test ◽  
Type I ◽  
Cross Sectional ◽  
Positive Expression ◽  
Rabbit Polyclonal Antibody ◽  
Histopathological Type

Background: Nasopharyngeal carcinoma (NPC) is a malignancy originated from nasopharyngeal epithelial cells. NPC therapy response could be predicted from histopathological type, but some patients with the same histopathological type, showed a different therapy response. Interleukin (IL)-10 expression is expected to be able to predict a better response of therapy in NPC patients. Purpose: To find out the correlation between IL-10 expression and histopathological type in NPC patients. Method: An analytic observational study with cross sectional approach towards 33 samples taken from the Oncology Polyclinic of Outpatient Unit of Otorhinolaryngology Head and Neck Surgery Department, Dr. Soetomo General Hospital. Formalin-fixed paraffin-embedded biopsy specimens were obtained. The IL-10 expression was studied with immunohistochemistry using rabbit polyclonal antibody Anti IL-10. Assessment of the staining used Allred scale. The Fisher exact test was utilized to determine the correlation of IL-10 expression and histopathological type of NPC, with p value = 0.05. Result: The result of IL-10 expression in NPC patients with histopathological WHO type I NPC obtained 1 sample (8.3%) with strong positive expression and 2 samples (9.5%) with weak positive expression. In patients with histopathological WHO type II NPC obtained 2 samples (16.7%) with strong positive expression and 12 samples (57.1%) with weak positive expression. In patients with histopathological WHO type III NPC obtained 9 samples (75%) with strong expression and 7 samples (33.3%) with weak positive expression. Conclusion: There was moderate positive correlation between IL-10 expression and histopathological type in NPC patients.Keywords: nasopharyngeal carcinoma, IL-10 expression, histopathological type ABSTRAK Latar belakang: Karsinoma nasofaring (KNF) adalah suatu keganasan yang berasal dari epitel nasofaring. Respon terapi KNF selama ini dapat dinilai berdasarkan tipe histopatologi, namun pada kenyataannya penderita KNF dengan tipe histopatologi sama dapat menunjukkan respon terapi berbeda. Pemeriksaan ekspresi interleukin (IL)-10 diharapkan dapat memberikan prediksi lebih baik mengenai respon terapi pada penderita KNF. Tujuan: Mengetahui hubungan antara ekspresi IL-10 dengan tipe histopatologi pada penderita KNF. Metode: Penelitian observasional analitik dengan pendekatan cross sectional pada 33 sampel yang diperoleh dari Poliklinik Onkologi Unit Rawat Jalan, Departemen THTBedah Kepala Leher, RSUD Dr Soetomo. Ekspresi IL-10 diperiksa dari blok parafin dengan teknik pemulasan imunohistokimia menggunakan rabbit polyclonal antibody Anti IL-10. Ekspresi IL-10 dinilai dengan menggunakan skala Allred. Uji Fisher exact digunakan untuk menentukan korelasi antara ekspresi IL-10 dan tipe histopatologi KNF, dengan p = 0,05. Hasil: Ekspresi IL-10 pada KNF WHO tipe I didapatkan ekspresi positif kuat 1 penderita (8,3%) dan ekspresi positif lemah 2 penderita (9,5%). Hasil ekspresi IL-10 pada KNF WHO tipe II didapatkan ekspresi positif kuat 2 penderita (16,7%) dan ekspresi positif lemah 12 penderita (57,1%). Hasil ekspresi IL-10 pada KNF WHO tipe III 9 penderita (75%) dengan ekspresi positif kuat dan 7 penderita (33,3%) dengan ekspresi positif lemah. Kesimpulan: Didapatkan korelasi positif sedang antara ekspresi IL-10 dan tipe histopatologi pada penderita KNF

scholarly journals Validation of commercially available sphingosine kinase 2 antibodies for use in immunoblotting, immunoprecipitation and immunofluorescence

F1000Research ◽  
2016 ◽  
Vol 5 ◽  
pp. 2825 ◽  
Author(s):  
Heidi A. Neubauer ◽  
Stuart M. Pitson
Keyword(s):  
Amino Acids ◽  
Cell Lines ◽  
Polyclonal Antibody ◽  
Human Cell ◽  
Sphingosine Kinase ◽  
Immunofluorescence Staining ◽  
Human Cell Lines ◽  
Sphingosine Kinase 2 ◽  
Hela Cell Lines ◽  
Rabbit Polyclonal Antibody

Sphingosine kinase 2 (SK2) is a ubiquitously expressed lipid kinase that has important, albeit complex and poorly understood, roles in regulating cell survival and cell death. In addition to being able to promote cell cycle arrest and apoptosis under certain conditions, it has recently been shown that SK2 can promote neoplastic transformation and tumorigenesis in vivo. Therefore, well validated and reliable tools are required to study and better understand the true functions of SK2. Here, we compare two commercially available SK2 antibodies: a rabbit polyclonal antibody from Proteintech that recognizes amino acids 266-618 of human SK2a, and a rabbit polyclonal antibody from ECM Biosciences that recognizes amino acids 36-52 of human SK2a. We examine the performance of these antibodies for use in immunoblotting, immunoprecipitation and immunofluorescence staining of endogenous SK2, using human HEK293 and HeLa cell lines, as well as mouse embryonic fibroblasts (MEFs). Furthermore, we assess the specificity of these antibodies to the target protein through the use of siRNA-mediated SK2 knockdown and SK2 knockout (Sphk2-/-) MEFs. Our results demonstrate that the Proteintech anti-SK2 antibody reproducibly displayed superior sensitivity and selectivity towards SK2 in immunoblot analyses, while the ECM Biosciences anti-SK2 antibody was reproducibly superior for SK2 immunoprecipitation and detection by immunofluorescence staining. Notably, both antibodies produced non-specific bands and staining in the MEFs, which was not observed with the human cell lines. Therefore, we conclude that the Proteintech SK2 antibody is a valuable reagent for use in immunoblot analyses, and the ECM Biosciences SK2 antibody is a useful tool for SK2 immunoprecipitation and immunofluorescence staining, at least in the human cell lines employed in this study.

Development and Characterization of Anti-Estradiol Polyclonal Antibody

2012 ◽  
Vol 461 ◽  
pp. 67-70 ◽  
Author(s):  
Chao Ying Li ◽  
Jin Qing Jiang
Keyword(s):  
Polyclonal Antibody ◽  
Dynamic Range ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Uv Absorbance ◽  
Standard Curve ◽  
Ic50 Value ◽  
New Zealand White Rabbits

This paper reports an indirect competitive enzyme-linked immunosorbent assay (icELISA) using polyclonal antibody (pAb) for estradiol (E2) residues. After derivation, E2 haptens were conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) through 1-Ethyl-3-(3-dimethylaminopropy) carbodiimide (EDC) method, and New Zealand white rabbits were immunized to produce anti-E2 pAb. The conjugation ratio of E2-BSA was proved to be 18.6:1 by an UV absorbance method. Based on the square matrix titration, an icELISA standard curve was developed. The dynamic range was from 0.16 to 128 ng/mL, with LOD and IC50 value of 0.08 ng/mL and 3.76 ng/mL, respectively. Except for a little cross-reactivity (16.2%) to estrone, this assay showed negligible cross-reactivity to other analogues tested. The results suggest that the produced anti-E2 pAb could be used to develop an icELISA method for the determination of E2 residues in animal-originally products.

scholarly journals Crosstalk Pathway between Trehalose Metabolism and Cytokinin Degradation for the Determination of the Number of Berries per Bunch in Grapes

Cells ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 2378
Author(s):  
Ayane Moriyama ◽  
Chiho Yamaguchi ◽  
Shinichi Enoki ◽  
Yoshinao Aoki ◽  
Shunji Suzuki
Keyword(s):  
Cultured Cells ◽  
Growing Season ◽  
Cytokinin Oxidase ◽  
Flower Formation ◽  
Bud Break ◽  
Flower Buds ◽  
New Information ◽  
Trehalose Metabolism ◽  
Cultivation Techniques

In grapes, the number of flowers per inflorescence determines the compactness of grape bunches. Grape cultivars with tight bunches and thin-skinned berries easily undergo berry splitting, especially in growing areas with heavy rainfall during the grapevine growing season, such as Japan. We report herein that grape cytokinin oxidase/dehydrogenase 5 (VvCKX5) determines the number of berries per inflorescence in grapes. The number of berries per bunch was inversely proportional to the VvCKX5 expression level in juvenile inflorescences among the cultivars tested. VvCKX5 overexpression drastically decreased the number of flower buds per inflorescence in Arabidopsis plants, suggesting that VvCKX5 might be one of the negative regulators of the number of flowers per inflorescence in grapes. Similarly, the overexpression of grape sister of ramose 3 (VvSRA), which encodes trehalose 6-phosphate phosphatase that catalyzes the conversion of trehalose-6-phosphate into trehalose, upregulated AtCKX7 expression in Arabidopsis plants, leading to a decrease in the number of flower buds per Arabidopsis inflorescence. VvCKX5 gene expression was upregulated in grapevine cultured cells and juvenile grape inflorescences treated with trehalose. Finally, injecting trehalose into swelling buds nearing bud break using a microsyringe decreased the number of berries per bunch by half. VvCKX5 overexpression in Arabidopsis plants had no effect on the number of secondary inflorescences from the main inflorescence, and similarly trehalose did not affect pedicel branching on grapevine inflorescences, suggesting that VvCKX5, as well as VvSRA-mediated trehalose metabolism, regulates flower formation but not inflorescence branching. These findings may provide new information on the crosstalk between VvSRA-mediated trehalose metabolism and VvCKX-mediated cytokinin degradation for determining the number of berries per bunch. Furthermore, this study is expected to contribute to the development of innovative cultivation techniques for loosening tight bunches.

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