Rat anti pdgfrβ

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Rat anti-PDGFRβ is a primary antibody that binds to the Platelet-Derived Growth Factor Receptor Beta (PDGFRβ) protein. It can be used in various immunoassay techniques to detect and study the expression of PDGFRβ in cellular and tissue samples.

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Lab products found in correlation

Rabbit anti zo 1, by Thermo Fisher Scientific (2 mentions) Frozen section compound, by Leica camera (1 mentions) Rabbit anti ng2 antibody, by Merck Group (1 mentions) Alexa fluor 546 secondary antibody, by Thermo Fisher Scientific (1 mentions) Biorevo, by Keyence (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions) Dm5500b microscope, by Leica camera (1 mentions) Horseradish peroxidase conjugated goat anti rabbit, by Abcam (1 mentions) Goat anti rat, by Thermo Fisher Scientific (1 mentions) Primary rabbit anti cd31, by Abcam (1 mentions) Guinea pig anti insulin, by Abcam (1 mentions) Image pro plus program, by Media Cybernetics (1 mentions) Rabbit anti cd31, by Abcam (1 mentions) Rabbit anti gfp, by Abcam (1 mentions) Alexa fluor 488 conjugated donkey anti rabbit, by Abcam (1 mentions) Dapi mounting medium, by Merck Group (1 mentions) Tcs sp8 confocal microscope, by Leica camera (1 mentions) Fluorophore conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Rabbit anti psmad3, by Abcam (1 mentions) Lsm 5 pascal microscope, by Zeiss (1 mentions)

9 protocols using rat anti pdgfrβ

Pericyte Labeling after Laser Injury

Cited 1 time

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Seven days after laser injury, eyes were enucleated and fixed in 4% para-formaldehyde for 1 h on ice, incubated in an increasing concentration of PBS/sucrose (5, 10, and 20%), and embedded in frozen section compound (Leica, Exton, PA, USA). After blocking, the sections were incubated with rat anti-PDGFR-β (1:100, Thermo Fisher Scientific) and rabbit anti-NG2 antibody (1:100, Millipore, USA) and subsequently Alexa Fluor 546 secondary antibodies (1:200, Molecular Probes, Waltham, MA, USA). The specimens were mounted in mounting medium, including DAPI (1:500, Vector Laboratories, Burlingame, CA, USA), and examined under a fluorescence microscope (BIOREVO, Keyence, Tokyo, Japan). The number of nucleus surrounded by NG-2 positive signals, corresponding to pericytes, are counted unbiasedly in five consecutive tissue sections, as previously described [17 (link),18 (link)].

Liu Y., Noda K., Murata M., Wu D., Kanda A, & Ishida S. (2020). Blockade of Platelet-Derived Growth Factor Signaling Inhibits Choroidal Neovascularization and Subretinal Fibrosis in Mice. Journal of Clinical Medicine, 9(7), 2242.

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Visualizing Pancreatic Islet Vasculature

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Integrity of pancreatic islet blood vessels was analyzed by light microscopy upon staining with antibodies to platelet endothelial cell adhesion molecule 1 (PECAM1/CD31) and pericyte marker, PDGFRβ. Briefly, pancreatic sections were stained with primary rabbit anti-CD31 (1:50; Abcam) or rat anti-PDGFRβ (1:100; Thermo Fisher) antibodies at 4°C overnight, followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit (1:2000; Abcam), or goat anti-rat (1:5000; Thermo Fisher) antibodies respectively, for 30 mins at room temperature. The sections were then incubated with the IHCWORLD substrate and 3-amino-9-ethylcarbazole (AEC) chromogen. Cell nuclei were counterstained with hematoxylin. The images were visualized and captured with a Leica DM5500 B microscope.

Ukah T.K., Cattin-Roy A.N., Davis G.E, & Zaghouani H. (2021). Formation of pancreatic β-cells from precursor cells contributes to the reversal of established type 1 diabetes. Cellular immunology, 364, 104360.

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Pancreatic Cell Analysis and Quantification

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Pancreatic sections were blocked with a PBS solution containing 1% BSA, 1% goat or donkey serum, and 0.2% Triton X-100 at RT for 1 hour. The sections were then incubated overnight at 4°C with primary antibodies, including rabbit anti-GFP (1:500; Abcam), guinea pig anti-insulin (1:300; Abcam), rabbit anti-CD31 (1:100; Abcam), rat anti-PDGFRβ (1:100; ThermoFisher) and eFluor 570-conjugated Ki-67 monoclonal Ab (1:200; Invitrogen). The slides were washed with three changes of 0.02% Triton X-100 in PBS and then stained for 1 hour at RT with the corresponding secondary antibodies, including Alexa Fluor 647–conjugated goat anti-guinea pig (1:300; Abcam), Alexa Fluor 488–conjugated donkey anti-rabbit, (1:500; Abcam) and Alexa Fluor 594–conjugated chicken anti-rat (1:400; Abcam). DAPI mounting medium (Sigma) was used for nuclear staining. The images were visualized and acquired with a Leica TCS SP8 confocal microscope. The number of β-cells and islets was scored with a computer-assisted Image Pro Plus program (Media Cybernetics, Silver Spring, MD). To determine the number of β-cell and islets per mouse, each pancreas was sectioned at every 150 μm, spanning both proximal and distal areas of the pancreas, and all the resulting sections were used for the analysis.

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Liver and Lung Fibrosis Analysis

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Paraffin-embedded sections were processed as described previously (17 (link)). Sections (5 µm) were stained with hematoxylin and eosin or with Sirius red. Pictures were taken from random fields from each section at a final magnification of ×10. Staining area was calculated by pixel counting with National Institutes of Health (NIH) ImageJ. For fluorescence microscopy, fixed livers and lungs were transferred to 30% sucrose in PBS overnight, embedded in optimum cutting temperature compound, and then cryosectioned at 5 µm. Cryosections were permeabilized and blocked with 0.3% Triton X-100 and 3% BSA in PBS. Sections were incubated with primary antibodies (rabbit anti-pSmad3, Epitomics, 1880-1, 1:100; rat anti-PDGFRβ, eBioscience, 14-1402, 1:100) overnight at 4 °C and then with fluorophore-conjugated secondary antibodies (Invitrogen). Confocal imaging was performed on a Zeiss LSM 5 Pascal microscope. pSmad immunofluorescence staining was quantified as described (32 (link)).

Reed N.I., Jo H., Chen C., Tsujino K., Arnold T.D., DeGrado W.F, & Sheppard D. (2015). The αvβ1 integrin plays a critical in vivo role in tissue fibrosis. Science translational medicine, 7(288), 288ra79.

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Immunostaining Analysis of Brain Injury

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Standard immunohistochemistry and immunofluorescence protocols were used to examine protein expression. Primary antibodies used in this study included: Rabbit anti-Laminin α5 (1:800, generated as described previously [42 (link)]), Rat anti-Laminin α2 (1:400, Sigma, USA), Goat anti-Laminin α4 (1:400, R&D, USA), Rat anti-CD31 (1:100, BD Biosciences, USA), Mouse anti-Claudin-5 (1:200, Invitrogen, USA), Rabbit anti-ZO-1 (1:400, Thermofisher, USA), Rat anti-PDGFRβ (1:200, eBioscience, USA), Rabbit anti-AQP4 (1:500, Millipore, USA), IB4–488 (1:200, Thermofischer, USA), Rat anti-Ly6G (1:200; Biolegend, USA), Rat anti-CD68 (1:200, Biolegend, USA), Rabbit anti-glial fibrillary acidic protein (GFAP, 1:400, Stem cell, USA), and Rabbit anti-Iba1 (1:500; Wako Inc, USA). The fluorescence intensity of immunoreactivity and the number of immune-positive cells were calculated using the ImageJ software (NIH, USA). For quantifications, 3–5 random fields around the hematoma per section, 5 non-consecutive sections per animal, and at least 3 animals were used.

Gautam J., Miner J.H, & Yao Y. (2019). Loss of endothelial laminin α5 exacerbates hemorrhagic brain injury. Translational stroke research, 10(6), 705-718.

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Multiparametric Kidney Histology Analysis

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Mice were euthanized under deep isoflurane anesthesia and immediately perfused via the left ventricle with ice-cold PBS for 1 min. Kidneys were harvested, cut in transversal sections, fixed in freshly prepared 4% paraformaldehyde and 5 μm cryosections (Tissue Tek, Sakura) were stained using the following primary antibodies: rat anti-F4/80 (Abcam, 1:1000), rat anti-CD31 (eBioscience, 1:600), rabbit anti-CD3 (eBioscience, 1:300), rabbit anti-laminin (Sigma, 1:2000), rat anti-PDGFRβ (eBioscience, 1:800), mouse anti-αSMA (Sigma, 1:2500), rat anti-Ly6g (BD Pharmingen, 1:500). Sections were subsequently incubated with corresponding Cy3- or Cy5-conjugated secondary antibodies (Jackson ImmunoResearch, 1:400). For PAS staining, kidney sections were fixed in formalin. Images were captured using Nikon C1 confocal and Zeiss Axio Observer Z1 microscope, respectively.

Hofmeister A., Thomaßen M.C., Markert S., Marquardt A., Preußner M., Rußwurm M., Schermuly R.T., Steinhoff U., Gröne H.J., Hoyer J., Humphreys B.D, & Grgic I. (2020). Development of a new macrophage-specific TRAP mouse (MacTRAP) and definition of the renal macrophage translational signature. Scientific Reports, 10, 7519.

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Targeted Adenoviral Delivery and Tissue Analysis

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Mice were administrated with 1×10 ¹¹ VP of triple targeted Ad via tail-vein injection. Three days post virus infection, mice were anesthetized and tissues (Liver, Lung, Pancreas, Spleen, Kidney, Small Bowel, Heart, Muscle and Brain) harvested for immunofluorescence staining. For frozen sections, organ slices were cryo-preserved in 30% sucrose in PBS at 4^oC overnight, embedded in NEG50 mounting medium (Thermo Fisher Scientific), and then frozen in a liquid nitrogen pre-chilled 2-methylbutane containing bucket. Sectioning of frozen organs was carried out using the CryoJane taping system (Leica Biosystems Inc). All frozen section slides were subject to immunofluorescence staining analysis in accordance with the standard techniques (8 (link)). Primary antibodies used in this study included hamster anti-CD31 (EMD Millipore), rat anti-endomucin (eBioscience), rat anti-PDGFRβ (eBioscience), rabbit anti-HSVtk (Dr. Summers’s lab) and Alexa Fluor 488 or 594-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories).

Lee M., Lu Z.H., Li J., Kashentseva E.A., Dmitriev I.P., Mendonca S.A, & Curiel D.T. (2020). Targeting tumor neoangiogenesis via targeted adenoviral vector to achieve effective cancer gene therapy for disseminated neoplastic disease. Molecular cancer therapeutics, 19(3), 966-971.

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Isolation of Myeloid Progenitors from Embryos

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Myeloid progenitors were isolated from E14.5–16.5 wild-type embryos by FACS following a modified procedure from that previously reported (Mukouyama et al., 2005 (link); Mukouyama et al., 2002 (link)). Enzyme-dissociated embryos were stained with the following fluorochrome-conjugated antibodies: rat anti-CD45 (eBioscience, clone 30-F11, 1:50) as a pan-hematopoietic cell marker; rat anti-F4/80 (eBioscience, clone BM8, 1:100) and anti-CD11b (e-Bioscience, clone M1/70, 1:25) as a monocyte/macrophage marker; rat anti-PDGFRβ (eBioscience, clone APB5, 1:100) as a pericyte marker. Cell viability was assessed using 7-aminoactinomycin D (Thermo Fisher Scientific, A1310). All sorting and analysis was performed with a FACS Aria II SORP instrument (BD Biosciences). Details of the culture experiment and staining procedure are available in the SUPPLEMENTAL EXPERIMENTAL PROCEDURES.

Yamazaki T., Nalbandian A., Uchida Y., Li W., Arnold T.D., Kubota Y., Yamamoto S., Ema M, & Mukouyama Y.S. (2017). TISSUE MYELOID PROGENITORS DIFFERENTIATE INTO PERICYTES THROUGH TGF-β SIGNALING IN DEVELOPING SKIN VASCULATURE. Cell reports, 18(12), 2991-3004.

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Immunofluorescent Staining and RNAscope Analysis of Pituitary Gland

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Pituitaries were fixed after dissection by immersion in 4% PFA at 4°C. Immunofluorescent stainings were performed on cryosections as previously described (56 (link)). The following primary antibodies were used: Goat anti-SOX2 (Biotechne, AF2018), rabbit anti-SOX9 (a gift from F. Poulat, IGH Montpellier, France), rat anti-GFP (Nacalai Tesque, 04404-84), chick anti-GFP (Invitrogen, A10262), rabbit anti-EDNRb (Abcam, 117529), goat anti-MSX1 (R&D Systems, AF5045), rabbit anti-ALDH1A2 (Abcam, 75674, after antigen retrieval), rabbit anti-Claudin3 (Abcam, ab15102), rat anti-PECAM (BD Pharmigen, 550274), rat anti-PDGFRβ (eBiosciences, 14-1402-81), rabbit anti-AQP3 (Alomone Labs, AQP-003, after antigen retrieval), rabbit anti-Runx1 (Sigma-Aldrich, HPA004176), rabbit anti-IRF9 (Cell Signalling Technology, D9I5H), goat anti-Met (R&D Systems, AF 527), and hormone antibodies anti-LH, GH, ACTH, and PRL from the National Hormone Peptide Program (A.F. Parlow, Torrance, USA). Slides were then incubated with Alexa Fluor secondary antibodies. Imaging of stained tissue sections was performed on a Leica SPE microscope while imaging of plated cells was done on an Olympus spinning disk microscope.
RNAscope was performed on cryosections following the manufacturer’s instructions using Hgf (#456511), Ednrb (#473801), and Pomc (#314081) probes.

Rizzoti K., Chakravarty P., Sheridan D, & Lovell-Badge R. (2023). SOX9-positive pituitary stem cells differ according to their position in the gland and maintenance of their progeny depends on context. Science Advances, 9(40), eadf6911.

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