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Dba fitc

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DBA-FITC is a fluorescent conjugate of Dolichos biflorus agglutinin (DBA), a plant lectin that binds specifically to N-acetylgalactosamine residues. DBA-FITC can be used to detect and visualize the distribution of N-acetylgalactosamine-containing glycoconjugates in biological samples.

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Lab products found in correlation

Lsm 780 confocal microscope, by Zeiss (3 mentions) Dolichos biflorus agglutinin fluorescein isothiocyanate, by Vector Laboratories (2 mentions) Afacsaria 3, by BD (2 mentions) Mouse fc block, by BD (2 mentions) Sytox red, by Thermo Fisher Scientific (2 mentions) Facsdiva software, by BD (2 mentions) Itgb1, by Abcam (1 mentions) Vinculin, by Merck Group (1 mentions) Cortactin, by Abcam (1 mentions) Itga2, by Abcam (1 mentions) Itga6, by Abcam (1 mentions) Nestin, by BD (1 mentions) Dba rhodamine, by Vector Laboratories (1 mentions) Axio scan z1 slide scanner, by Zeiss (1 mentions) Phalloidin tritc, by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Fluorescence microscope, by Olympus (1 mentions) Lightcycler 480, by Roche (1 mentions) Sybr green master mix, by Vazyme (1 mentions) Prism 5, by GraphPad (1 mentions)

Close spelling variants of this product

FITC-conjugated Dolichos biflorus agglutinin (DBA-FITC) (3 citations)

13 protocols using dba fitc

1

Mouse Toxoplasma Infection Virulence Evaluation

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Seven-week-old female ICR mice were infected with freshly egressed tachyzoites by intraperitoneal injection. Each strain or infection dose was tested by 5 mice. Symptoms and survival of infected mice were monitored daily for 30 days. At the conclusion of the virulence test, mice that survived the infections were euthanized and the number of Toxoplasma cysts in the brains was determined by Dolichos biflorus agglutinin-fluorescein isothiocyanate (DBA-FITC; Vector Laboratories, Burlingame, CA, USA) staining. All animal work was approved by the Ethical Committee of Huazhong Agricultural University (permit no. HZAUMO-2019-039).
Zhang J., Fan F., Zhang L, & Shen B. (2022). Nuclear Factor AP2X-4 Governs the Expression of Cell Cycle- and Life Stage-Regulated Genes and is Critical for Toxoplasma Growth. Microbiology Spectrum, 10(4), e00120-22.
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2

Isolation and Characterization of Liver Cholangiocytes

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Nonparenchymal liver cells were isolated from >P115
Alb-Cre+/−;Rbpjf/f;Hnf6f/fand
Rbpjf/f;Hnf6f/fmice as previously described51 (link). Cells were resuspended at 1 × 107 cells/mL
in DMEM/2% FBS and blocked with Mouse Fc Block (BD Biosciences) for 30 minutes.
Cells were incubated with fluorochrome-conjugated antibodies (Supplementary Table 2) and DBA-FITC
(Vector Laboratories) for 30 minutes, washed with cold DPBS 3 times and
resuspended in DMEM/2% FBS. Sytox Red (Thermo Fisher Scientific) was added to
label dead cells prior to sorting. Unstained and single-stained cells were used
for compensation. Specificity of DBA binding was verified with a GalNAc
(Sigma)-blocked control as previously described52 (link). Cells were analyzed and sorted on a
FACSAria III using FACSDiva software (BD Biosciences). From the
CD11b–CD31–CD45–population, EPCAM+DBA– cells were collected as
peripheral cholangiocytes and EPCAM+DBA+ as hilar
cholangiocytes. FlowJo (FlowJo, LLC) was used to analyze data and generate
charts. Cells were either sorted into DMEM/2% FBS, pelleted and snap frozen, or
sorted directly into extraction buffer for RNA purification.
Schaub J.R., Huppert K.A., Kurial S.N., Hsu B.Y., Cast A.E., Donnelly B., Karns R.A., Chen F., Rezvani M., Luu H.Y., Mattis A.N., Rougemont A.L., Rosenthal P., Huppert S.S, & Willenbring H. (2018). De novo formation of the biliary system by TGFβ-mediated hepatocyte transdifferentiation. Nature, 557(7704), 247-251.
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3

Comprehensive Tissue Analysis Protocol

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Histological and immunohistochemical analyses were performed as previously described2 (link). The following antibodies were used: Cdh1 (rat, Novex), Cortactin (rabbit, Abcam), GFP (goat, Abcam), GFP (mouse, Roche), ItgA2 (rabbit, Abcam), ItgA6 (rabbit, Abcam), ItgB1 (rabbit, Abcam), Ki67 (rat, Biolegend), Krt19 TROMA-III (rat, DSHB), MLC2 pSer19 (rabbit, NEB), Myosin (rabbit, Abcam), Nestin (mouse, BD transduction), Pdgfrβ (rabbit, NEB), PTK2 pTyr397 (rabbit, ThermoFisher), SMA (mouse, Agilent), SMA (mouse, Sigma), Tomato (rabbit, Rockland), Tomato (goat, Biorbyt), Vimentin (rabbit, NEB), Vinculin (mouse, Sigma-Aldrich). DBA-rhodamine and DBA-FITC were from Vectorlabs. F-actin was stained with Phalloidin-TRITC (Sigma-Aldrich) and nuclei with DAPI (Sigma-Aldrich). F-actin staining of LSL-KrasG12D; p53 F/F; Pdx1-Cre pancreata and wildtype littermate control pancreata was performed on cryosections. Samples were embedded fresh in OCT medium and after sectioning fixed in 5% NBF for 10 min. Slides were washed in 0.2% Triton X-100 in PBS for 10 min and incubated in FLASH blocking buffer for 30 min. Staining reagent incubations were performed as above. Fluorescent stainings were imaged on a Zeiss LSM 780 confocal microscope. Chromogenic DAB stainings were imaged on a Zeiss Axio Scan Z1 Slide Scanner.
Messal H.A., Alt S., Ferreira R.M., Gribben C., Wang V.M., Cotoi C., Salbreux G, & Behrens A. (2019). Tissue curvature and apicobasal mechanical tension imbalance instruct cancer morphogenesis. Nature, 566(7742), 126-130.
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4

Quantification of Toxoplasma Parasite Burden

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Groups of 5 mice were infected by i.p. injection of 1 × 102, 5 × 102, and 1 × 103 PruΔrop38 tachyzoites, and a group of 5 mice infected by i.p. injection of 1 × 103 Pru were used as control. Thirty days post-infection, the brains of mice infected with Pru and PruΔrop38 strains were harvested and homogenized in 2 mL volumes of sterile PBS using homogenizers. Cyst counts were performed on 200 μL of brain homogenates by DBA-FITC (Vector Laboratories, Burlingame, CA, USA) staining [23 (link)]. Cyst images were observed using a fluorescence microscope (Olympus Co., Tokyo, Japan). To quantify the brain parasite burden of mice, the number of parasites and DNA concentration in each brain tissue sample were detected by calculating the 529 gene of Toxoplasma and the 28S rRNA gene of the brain, respectively. Reaction conditions were 7 min incubation at 94 °C, followed by 45 cycles of 1 min at 94 °C, 1 min at 60 °C, 1 min at 72 °C and a final 10 min incubation at 72 °C using quantitative real-time PCR with the SYBR Green Master Mix (Vazyme Biotech, Nanjing, China) [24 (link)] (Supplementary Table S1). The DNA concentration was measured by spectrophotometry. Data were analyzed by a Roche LightCycler 480 (Roche, Basel, Switzerland).
Wu Y., Zhou Z., Ying Z., Xu Y., Liu J, & Liu Q. (2022). Deletion of Toxoplasma Rhoptry Protein 38 (PruΔrop38) as a Vaccine Candidate for Toxoplasmosis in a Murine Model. Biomedicines, 10(6), 1336.
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5

Isolation and Characterization of Liver Cholangiocytes

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Nonparenchymal liver cells were isolated from >P115
Alb-Cre+/−;Rbpjf/f;Hnf6f/fand
Rbpjf/f;Hnf6f/fmice as previously described51 (link). Cells were resuspended at 1 × 107 cells/mL
in DMEM/2% FBS and blocked with Mouse Fc Block (BD Biosciences) for 30 minutes.
Cells were incubated with fluorochrome-conjugated antibodies (Supplementary Table 2) and DBA-FITC
(Vector Laboratories) for 30 minutes, washed with cold DPBS 3 times and
resuspended in DMEM/2% FBS. Sytox Red (Thermo Fisher Scientific) was added to
label dead cells prior to sorting. Unstained and single-stained cells were used
for compensation. Specificity of DBA binding was verified with a GalNAc
(Sigma)-blocked control as previously described52 (link). Cells were analyzed and sorted on a
FACSAria III using FACSDiva software (BD Biosciences). From the
CD11b–CD31–CD45–population, EPCAM+DBA– cells were collected as
peripheral cholangiocytes and EPCAM+DBA+ as hilar
cholangiocytes. FlowJo (FlowJo, LLC) was used to analyze data and generate
charts. Cells were either sorted into DMEM/2% FBS, pelleted and snap frozen, or
sorted directly into extraction buffer for RNA purification.
Schaub J.R., Huppert K.A., Kurial S.N., Hsu B.Y., Cast A.E., Donnelly B., Karns R.A., Chen F., Rezvani M., Luu H.Y., Mattis A.N., Rougemont A.L., Rosenthal P., Huppert S.S, & Willenbring H. (2018). De novo formation of the biliary system by TGFβ-mediated hepatocyte transdifferentiation. Nature, 557(7704), 247-251.
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6

Cyst Quantification in Murine Toxoplasmosis

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Mice immunized with ME49Δα‐amy and naïve mice were infected with 104 tachyzoites of TgPIG‐WH1, and mice immunized but not TgPIG‐WH1 challenged were considered as a control group. After 30 days post‐infection, seropositive mice were euthanized and brain tissues of them were homogenized to measure the number of cysts by DBA‐FITC (Vector Laboratories, Burlingame, CA, USA) staining as described previously (Huskinson‐Mark et al., 1991).
Yang J., Yang C., Qian J., Li F., Zhao J, & Fang R. (2020). Toxoplasma gondii α‐amylase deletion mutant is a promising vaccine against acute and chronic toxoplasmosis. Microbial Biotechnology, 13(6), 2057-2069.
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7

Toxoplasma Infection in Mouse Model

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Freshly egressed tachyzoites of the ME49 strain and ME49ΔADSL strain were filtered with a 3.0 μm membrane filter and then harvested to intraperitoneally (ip) infect 8-week-old female ICR mice (10 mice per strain). Subsequently, the clinical symptoms of toxoplasmosis were observed, and mortality was recorded for 30 days. Blood samples were collected from the mouse tail vein at 30 days. After clot retraction, serum samples were collected and detected by enzyme-linked immunosorbent assay (ELISA) with T. gondii soluble antigens as coating antigen to confirm infection. Soluble Toxoplasma antigens were prepared by a previously described method [29 (link)]. Subsequently, brain cysts from seropositive mice were harvested, and DBA-FITC (Vector Laboratories, Burlingame, CA, USA) dyeing as previously described [30 (link)] was used to calculate the number of Toxoplasma cysts. The results of brain cyst number and cumulative mortality determination in the control and experimental group mice were graphed as Kaplan–Meier survival plots and then analyzed in Prism 5 (GraphPad Software Inc., La Jolla, CA, USA).
Wang L., Tang D., Yang C., Yang J, & Fang R. (2020). Toxoplasma gondiiADSL Knockout Provides Excellent Immune Protection against a Variety of Strains. Vaccines, 8(1), 16.
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8

Mouse Toxoplasma Infection Virulence Evaluation

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Seven-week-old female ICR mice were infected with freshly egressed tachyzoites by intraperitoneal injection. Each strain or infection dose was tested by 5 mice. Symptoms and survival of infected mice were monitored daily for 30 days. At the conclusion of the virulence test, mice that survived the infections were euthanized and the number of Toxoplasma cysts in the brains was determined by Dolichos biflorus agglutinin-fluorescein isothiocyanate (DBA-FITC; Vector Laboratories, Burlingame, CA, USA) staining. All animal work was approved by the Ethical Committee of Huazhong Agricultural University (permit no. HZAUMO-2019-039).
Zhang J., Fan F., Zhang L, & Shen B. (2022). Nuclear Factor AP2X-4 Governs the Expression of Cell Cycle- and Life Stage-Regulated Genes and is Critical for Toxoplasma Growth. Microbiology Spectrum, 10(4), e00120-22.
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9

Immunofluorescence Analysis of 3D Collagen Cultures

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Harvested tissues were fixed in 10% formalin and embedded in paraffin. Immunofluorescence analysis of tissue sections and cells seeded on coverslips was performed as shown previously [11 , 12 ]. Whole-mount immunofluorescence staining for 3D collagen cultures was performed, as shown previously [13 (link)]. The primary antibodies used in this study include PAF1 1:100 (Invitrogen Cat# PA5-115713), mouse PAF1 1:100 (In house developed) [3 (link)], YAP1 1:100 (Santa Cruz Cat# sc-271134), TEAD4 1:100 (Invitrogen Cat#PA5-41446), TEAD1 1:100 (Invitrogen Cat#PA5-106477), SOX9 1:200 (Sigma-Aldrich Cat# AMAB90795) and CK19 1:50 (DSHB Hybridoma Product TROMA-III). In some immunofluorescence experiments, conjugated lectins, PNA-Rhodamine 1:100 (Vector Cat# RL-1072), and DBA-FITC 1:100 (Vector Cat# FL-1031) were used to stain acinar and ductal cells. Fluorescent images were captured using an LSM 710 confocal microscope and analyzed using ZEN software.
Nimmakayala R.K., Ogunleye A.O., Parte S., Krishna Kumar N., Raut P., Varadharaj V., Perumal N.K., Nallasamy P., Rauth S., Cox J.L., Lele S.M., Batra S.K, & Ponnusamy M.P. (2022). PAF1 cooperates with YAP1 in metaplastic ducts to promote pancreatic cancer. Cell Death & Disease, 13(10), 839.
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10

Quantifying Toxoplasma Tissue Cysts

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Toxoplasma tachyzoites from wild type (WT) (ME49 Luc+ Δhxgprt), ME49 Δgra47, ME49 Δgra47 + H169R, and ME49 Δgra47 + GRA47 HA parasites, were obtained by lysing host HFFs through a 27-gauge needle. These tachyzoites were used to infect 6-week-old female CD-1 mice via intraperitoneal injection, with each mouse receiving 1,000 parasites. To assess parasite viability, a plaque assay was conducted immediately after infecting the mice. The mice were then observed daily and weighed every 2 days for a period of 30 days. At the end of this 30-day post-infection period, the mice were sacrificed, and their brains were collected to isolate tissue cysts. The mouse brain tissue was homogenized in PBS, and a fraction (1/10) of the homogenate was fixed with ice-cold methanol. The tissue cysts were subsequently stained using DBA-FITC (Vector Laboratories, #FL-1031–5) at a 1:500 dilution and quantified under a microscope.
Bitew M.A., Gaete P.S., Swale C., Maru P., Contreras J.E, & Saeij J.P. (2024). Two Toxoplasma gondii putative pore-forming proteins, GRA47 and GRA72, influence small molecule permeability of the parasitophorous vacuole. mBio, 15(3), e03081-23.
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