All sections were deparaffinized in xylene, hydrated in graded ethanol solutions, and immunostained with peroxidase conjugated secondary antibodies or fluorescent probes according to published protocols.[18 (link), 31 (link)] The following antibodies were used: rabbit anti-CD68 antibody (1/100, Abcam), rabbit anti-Fibronectin (FN) antibody (1/100, Sigma), rabbit anti-VEGF antibody (1/100, Santa Cruz Biotechnology), Mouse anti-Dentin Matrix Protein 1 (DMP1) antibody (1/2000, a gift from Dr. Chunlin Qin from Baylor College of Dentistry), Rabbit anti-Pigment epithelium-derived factor (PEDF) antibody (1/500, Millipore), Mouse anti-von Willebrand factor (vWF) antibody (1/100, Santa Cruz Biotechnology), rabbit anti-CD31 antibody (1/100, Abcam). Alizarin red staining to visualize calcium deposition was performed as per standard procedures. All fluorescently stained sections were imaged at the University of Illinois at Chicago Research Resource Center core imaging facility. Imaging was performed using a Zeiss LSM 710 confocal microscope equipped with Zen image analysis software and peroxidase stained sections were imaged using a Zeiss Axio-observer D1 microscope. All comparative fluorescence images were obtained using the same imaging conditions.
Pedf antibody
Manufactured by Merck Group
Sourced in United States
The PEDF antibody is a laboratory research tool used to detect and quantify the presence of the PEDF (Pigment Epithelium-Derived Factor) protein in biological samples. PEDF is a multifunctional secreted glycoprotein with important roles in various cellular processes. The PEDF antibody can be used in techniques such as Western blotting, immunohistochemistry, and ELISA to study PEDF expression and distribution.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 710 confocal microscope, by Zeiss (3 mentions)
Nanodrop 2000c, by Thermo Fisher Scientific (2 mentions)
Rabbit anti cd68 antibody, by Abcam (1 mentions)
Axio observer d1 microscope, by Zeiss (1 mentions)
Rabbit anti cd31 antibody, by Abcam (1 mentions)
Rabbit anti vegf antibody, by Santa Cruz Biotechnology (1 mentions)
Actin antibody, by Santa Cruz Biotechnology (1 mentions)
Nupage lds sample buffer, by Thermo Fisher Scientific (1 mentions)
Odyssey clx infrared imaging system, by LI COR (1 mentions)
Mouse vegf elisa kit, by Merck Group (1 mentions)
P iκbα, by Cell Signaling Technology (1 mentions)
Vegf c, by Cell Signaling Technology (1 mentions)
P ikkα, by Cell Signaling Technology (1 mentions)
P jnk, by Cell Signaling Technology (1 mentions)
β actin, by Merck Group (1 mentions)
3 protocols using pedf antibody
1
Immunohistochemical Analysis of Tissue Markers
2
Retinal Protein Extraction and Analysis
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Eyes were enucleated and placed in ice-cold 1 x PBS with protease inhibitor cocktail (pH 7.5; Roche Chemicals, Basel, Switzerland). The anterior segment, including the cornea, iris, ciliary body, and lens were removed. The posterior eyecups (2 eye cups per sample) consisting of the retina, choroid, and RPE were homogenized using a pestle pellet homogenizer in 100ul RIPA buffer. The buffer was then collected, pooled and centrifuged at 13,000 rpm. Protein concentration was then determined using the Nanodrop 2000c (Thermo Scientific) and supernatants were used for Western Blot and ELISA. Invitrogen NuPage LDS sample buffer were used for loading in a 12% gel for Western Blot analysis. Transfer was performed using the iBlot dry transfer equipment from Invitrogen. PEDF antibody (Millipore cat # 07–280) was used at 1:100 concentrations, actin antibody 1:50 (sc1615, Santa Cruz). LI-COR secondary antibodies were used for detection according to manufacturer’s protocol. Odyssey CLx Infrared imaging system (LI-COR Biosciences, USA) was used to visualize the bands. Mouse VEGF ELISA kit from Sigma (cat# RAB0509–1KT) was used to quantify VEGF levels according to the manufacturer’s protocol.
3
Antibody Procurement for Protein Analysis
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Antibodies for VEGF-C, P65, p-P65, IκBα, p-IκBα, IKKα, p-IKKα, P38, p-P38, JNK, and p-JNK were purchased from Cell Signaling Technology (Danvers, MA, USA), Antibodies for p-ERK (E4) and ERK 1 (K-23) from Santa Cruz Biotechnology (Santa Cruz, CA, USA), LYVE-1, histone 3, and D2-40 from Abcam (Cambridge, Massachusetts, USA) and β-actin from Sigma-Aldrich (Saint Louis, Missouri, USA), PEDF antibody from Millipore (Bedford, MA, USA), and IL-1β recombinant protein were from Sigma-Aldrich.
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