Mouse anti pecam 1

Manufactured by Santa Cruz Biotechnology

Sourced in United States, United Kingdom, Germany

Mouse anti-PECAM-1 is an antibody that specifically binds to the PECAM-1 (Platelet Endothelial Cell Adhesion Molecule-1) protein. PECAM-1 is a cell surface glycoprotein involved in cell-cell adhesion, particularly between endothelial cells. This antibody can be used to detect the presence and distribution of PECAM-1 in various cell and tissue samples.

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Lab products found in correlation

Fluorescence microscope, by Leica (2 mentions) Rabbit anti erk1 2, by Cell Signaling Technology (2 mentions) Goat anti vegfr2, by R&D Systems (2 mentions) Rabbit anti rfp, by Abcam (1 mentions) Rabbit anti ki67, by Cell Signaling Technology (1 mentions) Rabbit anti zo 1, by Thermo Fisher Scientific (1 mentions) Mouse anti occludin, by Thermo Fisher Scientific (1 mentions) Anti claudin 5, by Thermo Fisher Scientific (1 mentions) Mouse anti nf κb p65, by Santa Cruz Biotechnology (1 mentions) Donkey anti rabbit na934, by GE Healthcare (1 mentions) Mouse anti nqo 1, by Santa Cruz Biotechnology (1 mentions) Mouse anti ho1, by Santa Cruz Biotechnology (1 mentions) Rosiglitazone rsg a00183, by Adipogen (1 mentions) Rabbit anti pparγ, by Thermo Fisher Scientific (1 mentions) Mouse anti eea1, by BD (1 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Mouse anti α tubulin, by Santa Cruz Biotechnology (1 mentions) Mouse anti cd63, by Abcam (1 mentions) Mouse anti α tubulin clone dm1a, by Merck Group (1 mentions) Sheep anti tgn46, by Bio-Rad (1 mentions)

Spelling variants of this product

PECAM-1 (50 citations) Anti-PECAM-1 (16 citations) Mouse anti-human PECAM-1 (CD31; (3 citations)

7 protocols using mouse anti pecam 1

Immunofluorescence Staining for Angiogenesis

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Immunofluorescence staining was performed according to the protocol previously described [28 (link)]. The primary antibodies used were as follows: rabbit anti-RFP (1:200; Abcam Inc., USA), mouse anti-PECAM-1 (1:50; Santa Cruz Biotechnology, Germany), rabbit anti-Ki67 (1:200; Cell signaling technology, USA). Each section was washed with PBS and incubated with proper secondary antibodies at room temperature for 2 h the following day. 1%BSA was used as a control to confirm the specificity of the antibody and DAPI (1:30) was used to detect the nucleus. Images were acquired using a Leica fluorescence microscope (Leica, Germany). Ki67⁺ /CD31⁺ microvessels in the perifocal region was counted and quantified by an investigator who was blinded to the experimental groups.

Liu J., Li Q., Zhang K.S., Hu B., Niu X., Zhou S.M., Li S.G., Luo Y.P., Wang Y, & Deng Z.F. (2016). Downregulation of the Long Non-Coding RNA Meg3 Promotes Angiogenesis After Ischemic Brain Injury by Activating Notch Signaling. Molecular Neurobiology, 54(10), 8179-8190.

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Rosiglitazone Modulates Endothelial Barrier

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Rosiglitazone (RSG # A00183, MW: 357.4) was obtained from the Adipogen. Reagents and chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA) or Bio-Rad Laboratories (Hercules, CA, USA). The antibodies used in this study were purchased from the various sources: Rabbit anti-ZO-1 (#402200), mouse anti-occludin (#331500) and anti-claudin-5 (#4C3C2) from Life Technologies; rabbit anti-PPARγ (#331500) from Invitrogen; rabbit anti-Nrf2 (#NBP-1-32822) from Novus Biologicals. Mouse anti-PECAM-1 (#sc-376764), mouse anti-NQO-1 (#sc-376023), mouse anti-HO1 (#sc-390991) and mouse anti-NFκB-p65 (#sc-(F-6)-8008) from Santa Cruz Biotechnology. Donkey anti-rabbit (#NA934) and sheep anti-mouse (#NA931) HRP-linked secondary antibodies were obtained from GE Healthcare (Piscataway, NJ, USA).

Sivandzade F, & Cucullo L. (2019). Anti-Diabetic Countermeasures Against Tobacco Smoke-Dependent Cerebrovascular Toxicity: Use and Effect of Rosiglitazone. International Journal of Molecular Sciences, 20(17), 4225.

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HUVEC Isolation and Characterization

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Human umbilical vein endothelial cells (HUVECs) were isolated and cultured as previously described 30, 31. The following reagents were purchased: goat anti‐VEGFR2 (R&D Systems), rabbit anti‐phospho‐VEGFR2 (Y1175), rabbit antibodies to native and phosphorylated c‐Akt (S473), PLCγ1 (Y783) and p38 (T180/Y182), rabbit anti‐ERK1/2, mouse anti‐phospho‐ERK1/2 (T202, Y204), rabbit anti‐USP8 (Cell Signaling Technologies), rabbit anti‐TGN46, mouse anti‐EEA1 (BD Biosciences), mouse anti‐CD63 (AbCam), mouse anti‐α‐tubulin, mouse anti‐PECAM1, mouse anti‐transferrin receptor (TfR) (Santa Cruz Biotechnology), mouse FK2 anti‐ubiquitin (Affiniti Research Products), rabbit Apu2 anti‐K48‐linked ubiquitin (Millipore), rabbit Apu3 anti‐K63‐linked ubiquitin (Millipore), HRP‐conjugated secondary antibodies (PerBio Sciences), AlexaFluor‐conjugated secondary antibodies (Invitrogen), endothelial cell growth medium (ECGM) (PromoCell), non‐targeting and USP8 siGENOME SMARTpool siRNA duplexes (Dharmacon, GE Healthcare) and recombinant human VEGF‐A_165a (Genetech Inc.).

Smith G.A., Fearnley G.W., Abdul‐Zani I., Wheatcroft S.B., Tomlinson D.C., Harrison M.A, & Ponnambalam S. (2015). VEGFR2 Trafficking, Signaling and Proteolysis is Regulated by the Ubiquitin Isopeptidase USP8. Traffic (Copenhagen, Denmark), 17(1), 53-65.

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VEGFR Signaling Pathway Analysis

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Antibodies: goat-anti-VEGFR1 (#AF321), goat-anti-VEGFR2 (#AF357), rabbit-anti-phospho-VEGFR2-Y1214 (#AF1766) (R&D Systems, Minneapolis, MN, USA), rabbit-anti-Akt (#9272S), rabbit-anti-phospho-Akt (S473; #4060B), rabbit-anti-ERK1/2 (#9102S), mouse-anti-phospho-ERK1/2 (T202/Y204; #9106S), rabbit-anti-neuropilin 1 (NRP1; #3725S), rabbit-anti-phospho-VEGFR2-Y951 (#4991S), rabbit-anti-phospho-VEGFR2-Y1059 (#3817S), rabbit-anti-phospho-VEGFR2-Y1175 (#2478S; Cell Signaling Technology, Danvers, MA, USA), rabbit-anti-phospho-VEGFR2-Y1054 (Clone D1W; #04-894; Merck Millipore, Watford, UK), mouse-anti-α-tubulin (Clone DM1A; #T6199; Sigma Aldrich, Poole, UK), mouse-anti-PECAM-1 (CD31; #sc-65260; Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-ubiquitin (FK2; #14220; Caymen Chemical, MI, USA), mouse-anti-clathrin heavy chain antibody (X22; #ab2731; Abcam, Cambridge, UK), sheep-anti-TGN46 (#AHP500GT; AbD Serotec, Oxford, UK). Endothelial cell growth medium (ECGM) was from PromoCell (Heidelberg, Germany). Recombinant human VEGF-A₁₆₅ was from Genentech Inc. (San Francisco, CA, USA), both VEGF-A₁₂₁ and VEGF-A₁₄₅ was from Promocell.

Fearnley G.W., Smith G.A., Abdul-Zani I., Yuldasheva N., Mughal N.A., Homer-Vanniasinkam S., Kearney M.T., Zachary I.C., Tomlinson D.C., Harrison M.A., Wheatcroft S.B, & Ponnambalam S. (2016). VEGF-A isoforms program differential VEGFR2 signal transduction, trafficking and proteolysis. Biology Open, 5(5), 571-583.

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Rat Brain Immunofluorescence for Microvascular Density

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For rat brain immunofluorescence, rats were killed 48 h after MCAO. Brain samples (MCA cortex area) were fixed with 4% paraformaldehyde and then dehydrated sequentially in 20% and 30% sucrose solutions until the brain sank. Tissues were stored at −80°C after being embedded in optimal cutting temperature (OCT) compound. Consecutive coronal sections of 8 um thickness were incubated in 1% BSA with mouse anti-PECAM-1 (1 : 50; Santa Cruz Biotechnology, Heidelberg, Germany) or anti-CD34 (1 : 200; Abcam Inc., Cambridge, MA, USA) at 4°C overnight. Each section was washed with PBS and incubated with Alexa-488-conjugated goat anti-mouse secondary antibodies (1 : 400) or Alexa-594-conjugated donkey anti-rabbit secondary antibodies (1 : 400) at room temperature for 2 hours the following day. 1% BSA was used as a control to confirm the specificity of the antibody and DAPI (1 : 30) was used to detect the nucleus. To quantify microvascular density after MCAO, images of the ischemic cortex and the contralateral cortex were acquired using a Leica fluorescence microscope, the PECAM-1 positive cells or CD34 were positive cells counted in each image and the mean of the total positive cell counting was considered as microvascular density [14 (link), 15 (link)].

Liu J., Zhou X., Li Q., Zhou S.M., Hu B., Hu G.W., Niu X., Guo S.C., Wang Y, & Deng Z.F. (2017). Role of Phosphorylated HDAC4 in Stroke-Induced Angiogenesis. BioMed Research International, 2017, 2957538.

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Quantifying Tumor Necrosis and Angiogenesis

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Tumors were harvested, fixed in 4% paraformaldehyde for 24 h, embedded in paraffin and sectioned. All slide images were captured using an Aperio T3 Scanscope and quantification was performed by a blinded investigator using Aperio Imagescope Software (Aperio Technologies, Vista, CA).
Percent necrosis was determined from hematoxylin and eosin (H&E) stained sections. Unstained sections were stained with a monoclonal rabbit anti-mouse PECAM-1 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) and a polyclonal rabbit anti-human Ki67 (Novocastra Laboratories, Ltd., Newcastle, UK), to quantify, respectively, differences in blood vessel density and alterations in cellular proliferation. TUNEL staining was performed using a TdT In Situ Apoptosis Detection Kit – TACS Blue Label (Trevigen Inc., Gaithersburg, MD) per manufacturer’s instructions. Statistical significances of differences were calculated using the Student’s t-test, two-tailed.

Peters D.E., Hoover B., Cloud L.G., Liu S., Molinolo A.A., Leppla S.H, & Bugge T.H. (2014). Comparative toxicity and efficacy of engineered anthrax lethal toxin variants with broad anti-tumor activities. Toxicology and applied pharmacology, 279(2), 220-229.

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Tumor Angiogenesis Immunofluorescence Imaging

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For immunofluorescence analysis paraffin embedded tumor sections were stained with anti-VEGFR2 (Santa Cruz Biotechnologies), antihuman VWF (DakoCytomation) and anti-mouse PECAM1 (Santa Cruz Biotechnologies) antibodies followed by incubation with AlexaFluor 594-conjugated or AlexaFluor 488-conjugated secondary antibodies. Nuclei were counterstained with 4',6-diamidino,2-phenylindole (DAPI, Sigma). Tumor slices were analyzed using Confocal LSM510 microscope equipped with Plan-Neofluar 20 × /0.5 NA and Plan-Apochromat 63 × /1.4 NA oil objectives (Carl Zeiss). Z-stack images were elaborated using AxioVision Inside4D module (Carl Zeiss).

Corsini M., Ravelli C., Grillo E., Dell'Era P., Presta M, & Mitola S. (2021). Simultaneously characterization of tumoral angiogenesis and vasculogenesis in stem cell-derived teratomas. Experimental cell research, 400(2).

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