Rat anti pecam1

Manufactured by BD

Rat anti-PECAM1 is a monoclonal antibody that binds to the Platelet Endothelial Cell Adhesion Molecule 1 (PECAM1), also known as CD31. PECAM1 is a cell adhesion molecule expressed on the surface of endothelial cells, platelets, and certain immune cells. It plays a role in cell-cell interactions and the regulation of vascular permeability.

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Lab products found in correlation

Goat anti endomucin, by R&D Systems (3 mentions) Alexa fluor 568 anti goat igg, by Thermo Fisher Scientific (2 mentions) Alexa fluor 488 donkey anti rat igg, by Thermo Fisher Scientific (2 mentions) Alexa fluor 647 donkey anti rabbit igg, by Thermo Fisher Scientific (2 mentions) Alexa fluor 488 donkey anti mouse igg, by Thermo Fisher Scientific (2 mentions) Goat anti pdx1, by Abcam (2 mentions) Rat anti e cadherin, by R&D Systems (2 mentions) Rabbit anti pax8, by Proteintech (2 mentions) Goat anti hnf4α, by Santa Cruz Biotechnology (2 mentions) Goat anti alb, by Fortis Life Sciences (2 mentions) Goat anti foxa2, by Santa Cruz Biotechnology (2 mentions) Streptavidin alexa 488, by Thermo Fisher Scientific (1 mentions) Biotin conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Confocal microscope, by Leica (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Normal goat serum (ngs), by Merck Group (1 mentions) Rabbit anti laminin, by Merck Group (1 mentions) Cy3 or cy5, by Jackson ImmunoResearch (1 mentions) Rabbit anti eomes, by Abcam (1 mentions) Mouse anti α smooth muscle actin cy3, by Merck Group (1 mentions)

Close spelling variants of this product

Anti-PECAM1 (14 citations) Rat anti-CD31/PECAM1 (2 citations)

10 protocols using rat anti pecam1

Whole-Mount Immunohistochemistry for Tissue Analysis

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Whole-mount immunohistochemistry was performed using a modification of
the protocol from Ahnfelt-Rønne et al.
(2007) as previously described (Havrilak and Shannon, 2015b (link)). Optical sections were captured on a
Nikon AiRsi inverted laser microscope, and 3D images were created by a composite
of z-stacks using Bitplane Imaris software to process the fluorescent images. To
calculate the volume of tissue stained for ALB, we created isosurfaces using
smoothing thresholds and intensity values that allowed filtering out antibody
aggregates or any non-specific staining.
The primary antibodies used were: rabbit anti-NKX2-1 (1:3000, Seven
Hills Bioreagents), guinea pig anti-NKX2-1 (1:500, Seven Hills Bioreagents),
goat anti-endomucin (1:500, R & D Systems), rat anti-E-cadherin (1:2000,
R & D Systems), rabbit anti-PAX8 (1:500, Protein Tech), rat anti-PECAM-1
(1:500, BD Pharmigen), goat anti-HNF4α (1:200, Santa Cruz), goat
anti-PDX1 (1:5000, Abcam), goat anti-ALB (1:1000, Bethyl Laboratories), goat
anti-FOXA2 (1:500, Santa Cruz), and rat anti-LIV2 (1:200, MBL). Secondary
antibodies used were: Alexa Fluor 647 donkey anti-rabbit IgG, Alexa Fluor 568
anti-goat IgG, Alexa Fluor 488 donkey anti-rat IgG, Alexa Fluor 488 donkey
antimouse IgG (1:500, all from Life Technologies).

Havrilak J.A., Melton K.R, & Shannon J.M. (2017). Endothelial cells are not required for specification of respiratory progenitors. Developmental biology, 427(1), 93-105.

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Whole-Mount Immunohistochemistry for Tissue Analysis

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Havrilak J.A., Melton K.R, & Shannon J.M. (2017). Endothelial cells are not required for specification of respiratory progenitors. Developmental biology, 427(1), 93-105.

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Immunohistochemical Staining of OBSCs

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OBSCs were fixed for 20 min in 4% PFA in phosphate buffered saline (PBS), washed three times in PBS, then blocked for 1 h in PBS supplemented with 0.5% Triton X-100 and 3% normal Goat Serum (Sigma G9023). Slices were incubated at 4 °C in primary antibody (diluted in blocking solution) overnight. In order to detect PDGFRβ, heat-mediated antigen retrieval was performed in 10 mM citrate buffer (pH 6.0) for 40 min at 80 °C prior to primary antibody incubation. Slices were washed a further three times in 0.5% Triton-X100 in PBS (PBS-TX) then incubated with secondary antibodies (Life Technologies and Jackson) (1:500 dilution in blocking solution for 2 h at 4 °C). Three final PBS-TX washes were conducted before slices were mounted on slides and images captured using a Leica Confocal Microscope. Primary antibodies used: rabbit anti-PDGFRβ (28E1) (1:200, Cell Signalling, Cat. No: 3169S), rat anti-PECAM-1 (1:400, BD, Cat. No: 550274), rabbit anti-laminin (1:200, Sigma, Cat. No: L9393), Mouse MOAB-2 (pan Aβ) (1:1000, Merck-Millipore, Cat. No: MABN254) Rabbit Ki67 (1:1000, Abcam, Cat. No: ab15580) secondary staining was conducted using species-specific fluorophore-conjugated (Streptavidin Alexa 488, Molecular Probes; Cy3 or Cy5, Jackson,) or biotin-conjugated secondary antibodies (Jackson). DAPI (1 μg/mL, Sigma) was used to counterstain nuclei.

Durrant C.S., Ruscher K., Sheppard O., Coleman M.P, & Özen I. (2020). Beta secretase 1-dependent amyloid precursor protein processing promotes excessive vascular sprouting through NOTCH3 signalling. Cell Death & Disease, 11(2), 98.

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Embryo Fixation, Staining, and Genotyping

Cited 2 times

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Embryos were fixed, stained, imaged, and recovered for genotyping as previously described [68] (link). Primary and secondary antibody sources were described previously [3] (link), [50] (link), and also included rabbit anti-EOMES (Abcam), rabbit anti-DAB2 (Santa Cruz Biotech), rabbit anti-LAMA1 (Sigma), and rat anti-PECAM1 (BD Biosciences).

Wicklow E., Blij S., Frum T., Hirate Y., Lang R.A., Sasaki H, & Ralston A. (2014). HIPPO Pathway Members Restrict SOX2 to the Inner Cell Mass Where It Promotes ICM Fates in the Mouse Blastocyst. PLoS Genetics, 10(10), e1004618.

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Immunostaining Protocol for Mesenteric Vascular Structures

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Mesenteries were fixed in 4% paraformaldehyde at room temperature for 2 h and stained as previously described (Stanczuk et al., 2015 (link)). The following primary antibodies were used: mouse anti-α-smooth muscle actin-Cy3 (Sigma-Aldrich, C6198, 1:250), rat anti-endomucin (V.7C7) (Santa Cruz, SC-65495, 1:200), rat anti-CD41-FITC (eBioscience, 11-0411-81, 1:50), rabbit anti-collagen IV (Bio-Rad, 2150-1470, 1:500), chicken anti-GFP (Abcam, ab13970, 1:200), goat anti-neuropilin 2 (R&D Systems, AF567, 1:200), hamster anti-PECAM1 (Millipore, MAB1398Z, 1:1000), rat anti-PECAM1 (BD Pharmingen, 553370, 1:1000), rat anti-PECAM1-AF594 (BioLegend, 102520, 1:100), rabbit anti-human PROX1 (Stanczuk et al., 2015 (link); 1:200), rat anti-TER-119 (eBioscience, 145921, 1:200), rat anti-TER-119-AF647 (BioLegend, 116218, 1:50), goat anti-VE-cadherin (Santa Cruz, SC-6458, 1:200) and anti-podoplanin (Developmental Studies Hybridoma Bank at the University of Iowa, clone 8.1.1, 1:800). Autofluorescence signal at 550-600 nm wavelength was used for visualization of RBCs. Secondary antibodies conjugated to AF488, AF594, AF647 or Cy3 were from Jackson ImmunoResearch, and all were used at 1:300.

Zhang Y., Daubel N., Stritt S, & Mäkinen T. (2018). Transient loss of venous integrity during developmental vascular remodeling leads to red blood cell extravasation and clearance by lymphatic vessels. Development (Cambridge, England), 145(3), dev156745.

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Immunofluorescence Analysis of Lymphatic Markers

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The following primary antibodies were used: rabbit anti-β-gal (MP Biomedicals), rabbit anti-Prox1 (AngioBio and ProteinTech Group), goat anti-Prox1 (R&D Systems), hamster anti-podoplanin (Hybridoma Bank, Developmental Studies, University of Iowa), rat anti-PECAM1 (BD Pharmingen), goat anti-Vegfr3 (R&D Systems), rabbit anti-GFP (Molecular Probes), and chicken anti-GFP (Abcam). The following secondary antibodies were used: Alexa 488-conjugated donkey anti-rabbit (Molecular Probes), Alexa 488-conjugated donkey anti-guinea pig (Molecular Probes), Alexa 488-conjugated goat anti-hamster (Molecular Probes), Alexa 488-conjugated donkey anti-goat (Molecular Probes), Cy3-conjugated donkey anti-rabbit (Jackson ImmunoResearch Laboratories), Cy3-conjugated donkey anti-goat (Jackson ImmunoResearch Laboratories), and Cy5-conjugated donkey anti-rat (Jackson ImmunoResearch Laboratories).

Srinivasan R.S., Escobedo N., Yang Y., Interiano A., Dillard M.E., Finkelstein D., Mukatira S., Gil H.J., Nurmi H., Alitalo K, & Oliver G. (2014). The Prox1–Vegfr3 feedback loop maintains the identity and the number of lymphatic endothelial cell progenitors. Genes & Development, 28(19), 2175-2187.

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Spinal Cord Immunofluorescence Imaging

Cited 1 time

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After terminal anaesthesia by barbiturate overdose, mice were perfused transcardially with 4% paraformaldehyde and spinal cords processed for immunofluorescence as previously described^{60 (link),72 (link)}. Primary antibodies were: rat anti-Pecam1 (BD Biosciences 550274, 1:200). Secondary antibodies were: Alexa Fluor 555 Goat Anti Rat (1:200, Life Technologies, A21434). Immunofluorescence was imaged digitally on a slide scanner [Olympus VS-120 Slide scanner] or confocal microscope [Zeiss LSM880 + Airy fast module with ZEN 2 Black software (Zeiss, Oberkochen, Germany)]. Images were processed using ImageJ (NIH) or Imaris (Bitplane, version 9.0.0).

Squair J.W., Gautier M., Kathe C., Anderson M.A., James N.D., Hutson T.H., Hudelle R., Qaiser T., Matson K.J., Barraud Q., Levine A.J., La Manno G., Skinnider M.A, & Courtine G. (2021). Confronting false discoveries in single-cell differential expression. Nature Communications, 12, 5692.

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Allantois Explant Culture and Angiogenesis

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Briefly, Rbpj^f/f and Rbpj^−/− pregnant females were killed by cervical dislocation at E8.0. Allantois could be clearly identified at the end of PS in the exocoelom after wiping off the Reichert’s membrane. The floating allantoises were excised at the basal part, rinsed in cold CZB and culture medium three times, respectively, transferred to the 24-well Petri dish that has been coated with fibronectin, and then cultured for 24 h. The allantois explants were fixed in 4% PFA, permeabilized in 0.2% Triton X-100, and incubated with rat anti-PECAM1 (1:500, BD) overnight at 4 °C. Then they were washed in 0.1% BSA–PBS, incubated with the Cy3-labeled secondary antibody.

Lu J., Wu W., Xin Q., Zhou C., Wang J., Ni Z., Liu D., Xu Y., Yu Y., Yang N., Sun Y., He B., Kong S., Wang S., Wang C, & Wang H. (2019). Spatiotemporal coordination of trophoblast and allantoic Rbpj signaling directs normal placental morphogenesis. Cell Death & Disease, 10(6), 438.

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Tissue Fixation and Immunostaining Protocols

Cited 1 time

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Embryos harvested from timed matings were fixed by immersion and lungs from postnatal mice were inflation fixed using 4% paraformaldehyde (PFA) in phosphate-buffered saline (PBS). Following overnight immersion in 4% PFA/PBS, fixed tissue was processed according to standard protocols for paraffin or frozen embedding. Hematoxylin and eosin (H&E) staining, immunohistochemistry, and immunofluorescence were performed on tissue sections (5–10 μm) as previously described (Lange et al., 2009 (link)). Primary antibodies included guinea pig anti-Sox17 (Seven Hills Bioreagents), goat anti-endomucin (R&D Systems), rat anti-Pecam-1 (BD Pharmingen), GSL-IB4-biotin (Vector Labs), rat anti-CD34 (Abcam), goat anti-EphB4 (R&D Systems), and mouse anti-alpha smooth muscle actin (Sigma). Fluorophore-conjugated secondary antibodies included Alexa Fluor-488 and Alexa Fluor-594 (Jackson ImmunoResearch and Life Technologies). For fluorescent stains, sections were stained with DAPI and mounted with ProLong Gold anti-fade reagent following antibody labeling (Invitrogen). Bright-field images were obtained using a Zeiss Axio ImagerA2 microscope equipped with AxioVision Software. Fluorescent images were obtained using a Nikon A1Rsi inverted laser confocal microscope and analyzed using Imaris software (Bitplane Scientific Software).

Lange A.W., Haitchi H.M., LeCras T.D., Sridharan A., Xu Y., Wert S.E., James J., Udell N., Thurner P.J, & Whitsett J.A. (2014). Sox17 is required for normal pulmonary vascular morphogenesis. Developmental biology, 387(1), 109-120.

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Immunostaining and Imaging Yolk Sacs and BMDMs

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Yolk sacs were stained as described in Newton et al. [33 (link)]. Fixed tissues were stained with rat anti-PECAM-1 (BD, 550274) and rabbit anti-cleaved caspase-3 (CST, 9661). The following secondary antibodies were used: donkey anti-rabbit Cy3 (Jackson ImmunoResearch, 711-165-152) and donkey anti-rat Cy5 (Jackson ImmunoResearch, 712-175-153). Processed yolk sacs were mounted using ProLong Gold Antifade Mountant with DAPI (Invitrogen, P36392) and images were acquired using a LEICA SPE upright confocal microscope with 20x objective. On average about 50 optical sections were collected, representing about 50 μm deep volume, with 1.19 μm step size. The images shown are maximum intensity projections.
BMDMs were seeded at d5 on 4 chamber tissue culture treated glass slides (Falcon, 354104) and treated the following day. The cells were fixed in 4% PFA for 30 min at room temperature. Permeabilized in 0.25% Triton X-100 (w/v) and blocked in 5% BSA (w/v) and 0.05% Triton X-100 (w/v). Staining was performed with p65 (CST, 8242) for 2 h at room temperature followed by labeling with donkey anti-rabbit Cy3 (Jackson ImmunoResearch, 711-165-152) and 1:2000 Hoechst (H3569). The cells were mounted with ProLong Glass Antifade Mountant (P36980). Images were acquired using a LEICA SP8 inverted microscope with a 40x objective.

Kist M., Kőműves L.G., Goncharov T., Dugger D.L., Yu C., Roose-Girma M., Newton K., Webster J.D, & Vucic D. (2020). Impaired RIPK1 ubiquitination sensitizes mice to TNF toxicity and inflammatory cell death. Cell Death and Differentiation, 28(3), 985-1000.

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