Anti rabbit fitc

Manufactured by Abcam

Sourced in United States

Anti-rabbit FITC is a secondary antibody conjugated with the fluorescent dye FITC (Fluorescein Isothiocyanate). It is designed to detect and bind to primary antibodies raised in rabbit. This product is commonly used in immunohistochemistry, flow cytometry, and other fluorescence-based assays.

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Lab products found in correlation

Rabbit anti bsp, by Abcam (2 mentions) Sort1, by Abcam (1 mentions) Neuropilin 1 nrp 1, by R&D Systems (1 mentions) P75ntr, by Abcam (1 mentions) Rabbit igg, by Abcam (1 mentions) Dapi mounting media, by Vector Laboratories (1 mentions) Col2a1, by Abcam (1 mentions) Fix perm kit, by Thermo Fisher Scientific (1 mentions) Facsaria 3, by BD (1 mentions) Anti grp78 antibody, by Santa Cruz Biotechnology (1 mentions) Envisiontm, by Agilent Technologies (1 mentions) Dmi4000d microscope, by Leica camera (1 mentions) Anti mouse fitc, by Abcam (1 mentions) Trypsin edta, by Cambrex (1 mentions) Facsaria 2, by BD (1 mentions) Prolong gold antifade containing dapi, by Thermo Fisher Scientific (1 mentions) Rabbit anti gfp, by Abcam (1 mentions) Tissue tec o c t, by Sakura Finetek (1 mentions) Rabbit anti α sma antibody, by Proteintech (1 mentions) Mmp13, by Abcam (1 mentions)

9 protocols using anti rabbit fitc

Immunohistochemical Analysis of Spinal Cord and Submandibular Gland

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Serial 8 µm sections of spinal cord were analyzed for vesicular acetylcholine transporter (VAChT, diluted 1∶200) (Novus Biologicals, Littleton, CO, USA) immunoreactivity, as a marker of preganglionic sympathetic cholinergic neurons. Successive slides were used to analyze Sema3A immunoreactivity(diluted 1∶200). Bound antibodies were incubated with anti-goat IgG DyLight 594 (diluted 1∶500) (Bethyl, Montgomery, TX, USA) for VAChT and with anti-rabbit-FITC (diluted 1/500) (Abcam, Cambridge, UK) for Sema3A
Paraffin-embedded sections of SMG were incubated with primary antibodies neuropilin-1 (Nrp-1) (R&D Systems, Minneapolis, MN, USA) (diluted 1∶20), p75^NTR (diluted 1/500) (Abcam), Sort-1 (diluted 1/200) and TrkA (diluted 1/200). Bound antibodies were incubated with anti-rabbit-FITC. See data S1 for details.

Ezkurdia N., Raurell I., Rodríguez S., González A., Esteban R., Genescà J, & Martell M. (2014). Inhibition of Neuronal Apoptosis and Axonal Regression Ameliorates Sympathetic Atrophy and Hemodynamic Alterations in Portal Hypertensive Rats. PLoS ONE, 9(1), e84374.

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Immunostaining of Murine Small Intestine

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Small intestines were isolated from mice shortly after euthanization and flushed with PBS, the gut was then inverted onto a wooden skewer and placed in 10% neutral buffered formalin (NBF, Merck) solution for 4 h. The tissue was sliced longitudinally to remove from the skewer and tissue was then rolled up beginning at the ileum using the Swiss roll method as previously published⁷⁶. The tissue was stored in NBF solution O/N then placed into 70% ethanol prior to paraffin embedding. Thin sections of paraffin-embedded tissue were de-waxed and stained with anti-DCAMLK1 at 1:1000 (Abcam, UK) and secondary stained using anti-rabbit-FITC, control slides were incubated with rabbit IgG (Abcam, UK). Once stained, slides were mounted in DAPI mounting media (Vector Laboratories, UK) and imaged using the 10x objective on a Leica DiM8 microscope (Leica, Germany); images were analysed using ImageJ/Fiji.

Oftedal B.E., Maio S., Handel A.E., White M.P., Howie D., Davis S., Prevot N., Rota I.A., Deadman M.E., Kessler B.M., Fischer R., Trede N.S., Sezgin E., Maizels R.M, & Holländer G.A. (2021). The chaperonin CCT8 controls proteostasis essential for T cell maturation, selection, and function. Communications Biology, 4, 681.

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Chondrocyte Type II Collagen Expression

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Chondrocytes, treated with BC (1% w/v), CS (1% w/v), and untreated (CTR) samples (at a density of 1 × 10⁵ cells/sample) at the second passage of culture, were incubated with fluorescent‐labeled monoclonal antibody or respective isotype controls. The antibody used was anti‐collagen type II (COL2A1, AbCam). Secondary antibody was anti‐rabbit FITC (AbCam). For intracellular staining of COL2A1, cells were processed using the Fix & Perm Kit (Invitrogen) following the manufacturer's guidelines. The fluorescence associated to the cells was measured using FITC channel and calculated both as mean percentage of positivity for Type II collagen and as mean fluorescence intensity (MFI) for each sample. All data were acquired using FACS Aria III (BD) and analyzed using FCS version 3 software.

Stellavato A., Tirino V., de Novellis F., Della Vecchia A., Cinquegrani F., De Rosa M., Papaccio G, & Schiraldi C. (2016). Biotechnological Chondroitin a Novel Glycosamminoglycan With Remarkable Biological Function on Human Primary Chondrocytes. Journal of Cellular Biochemistry, 117(9), 2158-2169.

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Immunohistochemistry of GRP78 in Human Lung

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Formalin-fixed human lung tissue sections were de-paraffinized in a xylene series and rehydrated through a decreasing ethanol series for immunohistochemistry. The slides were pre-treated by microwave in citrate buffer (100 mM, pH 7.0) for 10 minutes, washed 3x with 1x PBS and 0.1% tween (TBS). Slides were incubated overnight at 4 °C in an anti-Grp78 antibody dilution (1:100) from Santa Cruz Biotechnology (Santa Cruz Biotechnology, Santa Cruz, CA, USA). EnVisionTM (DAKO, Bollschweil, Germany) was used for staining and detection, according to the manufacturer’s instructions. A Leica DMI 4000 D microscope was used to acquire images. For double immunofluorescence, tissue sections were incubated with primary antibodies specific to GRP78 (1:100) or SP-C (1:100) (surfactant protein C) (both: Santa Cruz Biotechnology, Santa Cruz, CA, USA). The secondary antibodies used were anti-goat Cy3 labelled to detect GRP78, and anti-rabbit FITC labelled to detect SP-C (Abcam, USA); both secondary antibodies were diluted 1:1000.

Nita I., Hostettler K., Tamo L., Medová M., Bombaci G., Zhong J., Allam R., Zimmer Y., Roth M., Geiser T, & Gazdhar A. (2017). Hepatocyte growth factor secreted by bone marrow stem cell reduce ER stress and improves repair in alveolar epithelial II cells. Scientific Reports, 7, 41901.

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Intracellular Protein Expression Analysis

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Cells were detached using trypsin-EDTA (200 mg/l EDTA, 500 mg/l trypsin; Cambrex, East Rutherford, NJ, USA, http://www.cambrex.com). Intracellular staining for OC, BSP, and OPN was performed using FIX & PERM Cell Fixation & Cell Permeabilization Kit (Life Technologies Laboratories, Monza, Italy, http://www.lifetechnologies.com) according to the manufacturer's procedure. At least 300,000 cells were incubated with indirect or direct fluorescent-conjugated antibodies. The primary antibodies used were: Phycoerythrin (PE)-conjugated anti-h-OC and Carboxyfluorescein (CFS)-conjugated anti-h-OPN (R&D Systems, Minneapolis, MN), rabbit anti-HDAC1 and mouse anti-HDAC2 (SantaCruz, Santa Cruz, CA, http://www.scbt.com.), and rabbit anti-BSP (Abcam, Cambridge, UK). The secondary antibodies were anti-rabbit FITC and anti-mouse FITC, all purchased from Abcam. For negative controls, cells were stained with an isotype control antibody. Labeled cells were analyzed by flow cytometry using a FACS Aria II (BD Biosciences, San Jose, CA) and all data analyzed with FCS Express version 3 software.

Paino F., la Noce M., Tirino V., Naddeo P., Desiderio V., Pirozzi G., De Rosa A., Laino L., Altucci L, & Papaccio G. (2014). Histone Deacetylase Inhibition with Valproic Acid Downregulates Osteocalcin Gene Expression in Human Dental Pulp Stem Cells and Osteoblasts: Evidence for HDAC2 Involvement. Stem Cells (Dayton, Ohio), 32(1), 279-289.

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Immunofluorescence Analysis of GFP Expression

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Skin and muscle from the site of vaccination were isolated and fixed in 4% paraformaldehyde and 10% sucrose in PBS at 4 °C for 30 min to overnight. Tissues were further embedded in Tissue-Tec^® O.C.T (Sakura Finetek, Torrance, CA, USA) above liquid nitrogen. Cryosections of 4 μm were cut from the frozen tissue blocks and mounted on poly-l-lysine-coated slides. Slides were dried and stored at −20 °C until use.
Slides were brought to RT and placed in PBS for 5 min to remove the OCT and stained with rabbit anti-GFP (Abcam, Cambridge, UK). Anti-rabbit FITC (Abcam) was used as a secondary antibody. Slides were mounted with Prolong Gold antifade containing DAPI (Invitrogen, Carlsbad, CA, USA). Staining was evaluated by fluorescence microscopy.

van de Wall S., Walczak M., van Rooij N., Hoogeboom B.N., Meijerhof T., Nijman H.W, & Daemen T. (2015). Tattoo Delivery of a Semliki Forest Virus-Based Vaccine Encoding Human Papillomavirus E6 and E7. Vaccines, 3(2), 221-238.

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Histones Induce Collagen I and α-SMA Expression

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LX2 cells were seeded in eight-well chamber slides and treated with or without 5 μg/mL histones for 6 d. The cells were fixed with 4% (w/v) paraformaldehyde for 10 min and permeabilized with PBS + 1% (v/v) Triton X-100 for 10 min. After blocking with 5% (w/v) dried milk + 2% (w/v) bovine serum albumin in PBS, the cells were incubated with sheep anti-human collagen I α1 antibody (1:200) or rabbit-anti-α-SMA antibody (1:200, Proteintech, Rosemont, IL, United States) at 4 °C overnight. After extensive washing, the cells were incubated with anti-sheep-fluorescein isothiocyanate (FITC, Abcam, 1:2000) or anti-rabbit-FITC (Abcam, 1:2000) at room temperature for 45 min. Following extensive washing, the slides were mounted on coverslips using mounting medium and antifade (Thermo Fisher Scientific, Renfrew, United Kingdom). The fluorescent images were visualized using an LSM-10 confocal microscope (excitation: 488 nm and emission: 512 nm).

Wang Z., Cheng Z.X., Abrams S.T., Lin Z.Q., Yates E., Yu Q., Yu W.P., Chen P.S., Toh C.H, & Wang G.Z. (2020). Extracellular histones stimulate collagen expression in vitro and promote liver fibrogenesis in a mouse model via the TLR4-MyD88 signaling pathway. World Journal of Gastroenterology, 26(47), 7513-7527.

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Immunofluorescence Analysis of Cellular Markers

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The cultured tissue blocks were embedded in paraffin and sectioned at the thickness of 4 µm. Samples were fixed with 4% paraformaldehyde for 15 min and washed in PBS at room temperature. Then, samples were permeabilized with 5% BSA (Calico LLC) and 0.1% Triton X-100 for 1 h at room temperature. Subsequently, samples were incubated the following primary antibodies: Rabbit vimentin (1 mg/ml; Abcam; cat. no. ab63379), CD90 (1:100; Abcam; cat. no. ab125215), IL-1β (1:150; Abcam; cat. no. 1743-1), p16 (1:200; Abcam; cat. no. 1712-1) and matrix metalloproteinase-13 (MMP-13; 1:200; Abcam; cat. no. ab9128) at 4°C overnight. Samples were then incubated with anti-rabbit FITC (1:200; Abcam; cat. no. ab27912) and anti-rabbit CY3 (1:200; Abcam; cat. no. ab6939) conjugated secondary antibody at room temperature for 2 h. After washing with PBS, the sections were sealed with fluorescent sealing tablets and observed under an Olympus fluorescent microscope at magnification, ×400 (Olympus Corporation) after counterstaining with DAPI (Nanjing KeyGen Biotech Co. Ltd.), which was used to counterstain the nuclei for 5 min at room temperature.

Zhang Y., Zhou S., Cai W., Han G., Li J., Chen M, & Li H. (2020). Hypoxia/reoxygenation activates the JNK pathway and accelerates synovial senescence. Molecular Medicine Reports, 22(1), 265-276.

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Evaluating DPSC Osteogenic Differentiation on bECM and Col-I Hydrogels

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DPSCs were cultured on bECM hydrogels (3, 4, 6, 8 mg/mL) and Col-I hydrogels (3, 4, 6, 8 mg/mL) for 3 weeks (n=24, Table S1). Subsequently, DPSCs/bECM hydrogel constructs were rinsed with PBS and fixed in 4% paraformaldehyde for 20 min at RT as described previously. [36] After washing three times with PBS, the cells were permeabilized with 0.2% Triton X-100 (Sigma) in PBS for 10 min at RT. The cells were then washed three times with PBS and blocked with 3% BSA for 30 min at RT. The DPSCs/bECM hydrogel constructs were subsequently incubated for 12 h with indirect or direct fluorescent-conjugated antibodies at 4°C. The primary antibodies used were FITCconjugated anti-OCN (R&D Systems), and rabbit anti-BSP (Abcam). The secondary antibody was anti-rabbit FITC purchased from Abcam. For negative controls, cells were stained with an isotype control antibody. The cell nuclei were stained with a 1 μg/mL solution of 4,6-diamidino-2-phenylindole (DAPI; Sigma) for 5 min. The cells were subsequently mounted with anti-fading medium (ProLong Antifade; Invitrogen) and observed by confocal microscopy (Leica; TCS SP5). The samples were not sectioned and all images (n>5 fields for sample) were displayed as maximum intensity z-projections.

Paduano F., Marrelli M., Alom N., Amer M., White L.J., Shakesheff K.M, & Tatullo M. (2017). Decellularized bone extracellular matrix and human dental pulp stem cells as a construct for bone regeneration. Journal of biomaterials science. Polymer edition, 28(8).

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