After the formalin-fixed paraffin-embedded specimens were deparaffinized and rehydrated, antigen retrieval was performed. Samples were blocked at 4 deg C overnight in 5% FBS/PBST, and then incubated for 4 hours at room temperature in Anti-VEGF antibody (Novus biologicals) (1:50) diluted with 5% FBS/PBST. Negative control of 5% FBS/PBST was also used.
Anti vegf antibody
Manufactured by Novus Biologicals
Sourced in United States
The Anti-VEGF antibody is a laboratory reagent used in various research applications. It functions by specifically binding to and neutralizing the vascular endothelial growth factor (VEGF), a key regulator of angiogenesis.
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Lab products found in correlation
Digital slide scanner, by 3DHISTECH (1 mentions)
Horseradish peroxidase conjugated streptavidin biotin complex, by Agilent Technologies (1 mentions)
Mayer s hematoxylin, by Agilent Technologies (1 mentions)
Pierce ecl plus reagent, by Thermo Fisher Scientific (1 mentions)
Molecular imager chemidoc xrs, by Bio-Rad (1 mentions)
Image labtm software, by Bio-Rad (1 mentions)
3 protocols using anti vegf antibody
1
VEGF Immunohistochemistry in FFPE Tissues
2
Retinal Degeneration Induced by H2O2
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To analyze the induction of retinal degeneration with a H 2 O 2 injection and to analyze VEGF expression, eyeball specimens for each group (n = 6) were collected and fixed in 10% neutral buffered formalin for histological examination. All samples were embedded in paraffin, and 5-μmthick sections were obtained for hematoxylin and eosin staining. To observe the degree of degeneration to evaluate VEGF expression, each section of the eyeball was stained with an anti-VEGF antibody (1:200; Novus Biologicals, Centennial, CO, USA) using immunohistochemistry (Supplementary Fig. 3). Reactions with a horseradish peroxidase-conjugated streptavidin-biotin complex (DAKO, Santa Clara, CA, USA) and 3,3-diaminobenzidine (EnVision Systems, Santa Clara, CA, USA) were performed to generate a chromatic signal. Mayer's hematoxylin (DAKO) was used as a counterstain. Morphometric images of whole sections in the eyeball were viewed using a digital slide scanner (3DHISTECH, Ltd, Budapest, Hungary).
3
VEGF Protein Analysis in Irradiated HUVECs
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HUVECs were harvested at 96 h post irradiation. A total of 40 μg cell protein was separated by a 10% SDS-PAGE gel, which was then transferred onto a PVDF membrane and probed with an anti-VEGF antibody (1:500, NOVUS, USA). GAPDH was employed as an internal standard, and the Pierce ECL Plus reagent (1:1000, Thermo, USA) was used to display the protein bands. Images were acquired by Molecular Imager ChemiDocTM XRS+ with Image Lab TM software (Bio-Rad, USA).
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