To evaluate the inflammation and epidermal regeneration in the wound area, tissues containing the wound site and their surrounding healthy skin were collected. The tissue samples were fixed in 4% (v/v) paraformaldehyde for 1 h right after sacrifice before embedded in paraffin. The samples were cross sectioned to slices (4 μm thickness) and then stained by Hematoxylin-Eosin (H&E). All slides were scanned and analyzed by a Digital Slide Scanner (KFBIO, Ningbo). The regenerated skins from the wound site were also excised for IF staining with Anti-VEGFA antibody (proteintech) and TNFA Ab (proteintech), respectively. CoraLite488-conjugated Affinipure Goat Anti-Mouse IgG(H + L) (proteintech) and CoraLite594-conjugated Goat Anti-Rabbit IgG(H + L) (proteintech) were used as the secondary antibody to reveal VEGFA and TNFA expression. The nuclei were stained with 4′,6-diamidino-2-phenylindole. Slides were observed under an upright fluorescence microscope (BX53, Olympus).
Anti vegfa antibody
Manufactured by Proteintech
Sourced in United Kingdom
The Anti-VEGFA antibody is a laboratory reagent used to detect and quantify the presence of Vascular Endothelial Growth Factor A (VEGFA) in biological samples. It is a highly specific and sensitive tool for researchers studying angiogenesis, tumor growth, and various vascular-related processes.
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Lab products found in correlation
Coralite488 conjugated affinipure goat anti mouse igg h l, by Proteintech (1 mentions)
Coralite594 conjugated goat anti rabbit igg h l, by Proteintech (1 mentions)
Anti ki67 antibody, by Proteintech (1 mentions)
Anti cd163 antibody, by Proteintech (1 mentions)
Anti cd8 alpha antibody, by Abcam (1 mentions)
Moticeasyscan, by Motic (1 mentions)
Anti e cadherin antibody, by Proteintech (1 mentions)
Anti n cadherin antibody, by Proteintech (1 mentions)
Anti vimentin antibody, by Proteintech (1 mentions)
Anti cd31 antibody, by Proteintech (1 mentions)
4 protocols using anti vegfa antibody
1
Skin Wound Healing and Inflammation Analysis
2
Protein Expression Analysis by Western Blot and IHC
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Western blotting and IHC analysis were performed as described previously (Cao et al., 2018 (link)). The antibodies used were as follows: anti-FNDC3A antibody (Abcam, Cambridge, United Kingdom), anti-VEGFA antibody (Proteintech, Wuhan, China), anti-OPN3 antibody (Affinity Biosciences, Cincinnati, United States), and anti-CPE antibody (Proteintech, Wuhan, China). The IHC quantization analysis was calculated by ImageJ software and statistically analyzed in three random fields. Data are shown as mean ± SEM.
3
Immunohistochemical Analysis of Cancer Biomarkers
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Thirty-six representative samples from C1 (N = 18) and C2 (N = 18) were selected for tissue microarray (TMA) construction, the TMA is prepared as previously described43 . For immunohistochemistry staining, the sections were stained using anti-Vimentin antibody (Proteintech, 1:2000), anti-Ki67 antibody (Proteintech, 1:5000), anti-Cyclin D antibody (Proteintech, 1:1500), anti- E-cadherin antibody (Proteintech, 1:2000), anti- N-cadherin antibody (Proteintech, 1:200), anti-ADH1A antibody (Abcam, 1:500), anti-CYP3A4 antibody (Proteintech, 1:200), anti-CD34 antibody (Proteintech, 1:1000), anti-VEGFA antibody (Proteintech, 1:200), anti-CD8-alpha antibody (Abcam, 1:500), anti-CD163 antibody (Proteintech, 1:1000). Images were captured by MoticEasyScan (Motic).
4
Immunohistochemical Analysis of SACC Tumors
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Specimens of surgical tumour tissues from SACC patients were fixed with 4% formalin, paraffin embedded and sectioned (4 μm). The tissue sections were then deparaffinized and dehydrated followed by incubation in 3% hydrogen peroxide for 10 minutes. Slides were stained with primary antibodies at 4°C overnight after blocking with appropriate serum. Corresponding secondary antibodies were used for 1h at room temperature. Targeted molecules were detected following DAB staining for immunohistochemistry. Tissues were sectioned for CD31 immunohistochemical staining, followed by PAS staining. Slides were finally counterstained with haematoxylin. Two independent investigators blinded to sample identify the anti‐VEGFA antibody (1:200 dilutions, ProteinTech) and anti‐CD31 antibody (1:100 dilutions, ProteinTech) primary antibody, respectively. PBS was used as the primary antibody for the negative controls.
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