Protein extraction and immunoblotting were performed as described in a previous study.30 Briefly, RIPA lysis buffer was added to tissue or cell samples to obtain total protein. Then, 5× SDS buffer was added to quantify the protein samples at 99°C for 10 min to desaturate the protein. The proteins were separated by SDS–PAGE electrophoresis, transferred to PVDF membranes, blocked with 3% nonfat dry milk (PBST) for 1 h, incubated with a VEGFA antibody (AF5131, Affinity Biosciences) overnight at 4°C, washed three times with PBST, incubated with a secondary antibody for 90 min at room temperature and washed three times with PBST. Finally, chemiluminescence imaging was performed to detect protein expression levels on the membranes.
Vegfa antibody
Manufactured by Affinity Biosciences
Sourced in China
The VEGFA antibody is a laboratory reagent used in research applications. It is a monoclonal antibody that specifically binds to and detects the VEGFA protein. VEGFA is a key regulator of angiogenesis, the process of new blood vessel formation. The VEGFA antibody can be used in various experimental techniques, such as Western blotting, immunohistochemistry, and enzyme-linked immunosorbent assay (ELISA), to study the expression and function of VEGFA in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Emodin, by Chengdu Must Bio-Technology (1 mentions)
Alexa fluor 555 labeled donkey anti mouse igg h l, by Beyotime (1 mentions)
2 3 5 triphenyltetrazolium chloride, by Merck Group (1 mentions)
Anti gapdh, by Affinity Biosciences (1 mentions)
Il 6 antibody, by Abcam (1 mentions)
Talent qpcr premix sybr green, by Tiangen Biotech (1 mentions)
Anti occludin, by Abcam (1 mentions)
Anti mmp2, by Beyotime (1 mentions)
Fastking gdna dispelling rt supermix, by Tiangen Biotech (1 mentions)
Antifade mounting medium, by Beyotime (1 mentions)
Triton x 100, by Beyotime (1 mentions)
Anti mmp9, by Abcam (1 mentions)
Rna easy fast tissue cell kit, by Tiangen Biotech (1 mentions)
Hrp labeled goat anti rabbit igg h l, by Beyotime (1 mentions)
Neutral balsam, by Solarbio (1 mentions)
Azocasein, by Merck Group (1 mentions)
β actin 13e5 rabbit mab, by Cell Signaling Technology (1 mentions)
Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (1 mentions)
Bca protein assay kit, by Solarbio (1 mentions)
3 protocols using vegfa antibody
1
Protein Extraction and Immunoblotting
2
Emodin Regulates Neuroinflammation Pathways
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The following
materials and reagents were used in the study: Emodin (no.: A0044,
Chengdu Must Biotechnology Co., Ltd., China, purity≥98%); anti-MMP9
(no.: Ab38898, Abcam, U.K.); anti-NF-κB p65 (no.: #8242, CST,
USA); anti-IKKβ (no.: #2678, CST, USA); 2,3,5-triphenyl tetrazolium
chloride (no.: T8877-25G, Sigma, USA); hematoxylin and eosin (HE)
staining kit (no.:BP-DL017, SenBeiJia Biological Technology Co., Ltd.
China); HRP-labeled goat anti-rabbit IgG (H + L) (no.: A0208, Beyotime
Biotechnology, China); anti-MMP2 (no.: Ab181286), anti-occludin (no.:
Ab216327), anti-extracellular signal-regulated kinase [ERK1/2 (no.:
Ab184699)], and p-anti-ERK1/2 (no.: Ab214036)—Abcam, U.K.;
IL-6 antibody (no.: DF6087), HIF1A antibody (no./AF1009), claudin-5
(no./AF5216), and VEGFA antibody (no.: DF7470)—Affinity, China;
Alexa Fluor 555-labeled donkey anti-mouse IgG(H + L) (A0460), Triton
X-100 (no.: ST795), and antifade mounting medium (no.: P0126)—Beyotime
Biotechnology, China; neutral balsam (no.: G8590, Beijing Solarbio
Science & Technology Co., Ltd. China); RNA Easy Fast Tissue/Cell
Kit (no.: DP451), FastKing gDNA Dispelling RT SuperMix (no.: KR118),
and Talent qPCR PreMix (SYBR Green) (no.: FP209)—Tiangen Biotech
(Beijing) Co., Ltd, China; anti-GAPDH (no: AF7021, Affinity); and
BCA Protein Assay Kit (no: P0012S, Beyotime Biotechnology, China).
materials and reagents were used in the study: Emodin (no.: A0044,
Chengdu Must Biotechnology Co., Ltd., China, purity≥98%); anti-MMP9
(no.: Ab38898, Abcam, U.K.); anti-NF-κB p65 (no.: #8242, CST,
USA); anti-IKKβ (no.: #2678, CST, USA); 2,3,5-triphenyl tetrazolium
chloride (no.: T8877-25G, Sigma, USA); hematoxylin and eosin (HE)
staining kit (no.:BP-DL017, SenBeiJia Biological Technology Co., Ltd.
China); HRP-labeled goat anti-rabbit IgG (H + L) (no.: A0208, Beyotime
Biotechnology, China); anti-MMP2 (no.: Ab181286), anti-occludin (no.:
Ab216327), anti-extracellular signal-regulated kinase [ERK1/2 (no.:
Ab184699)], and p-anti-ERK1/2 (no.: Ab214036)—Abcam, U.K.;
IL-6 antibody (no.: DF6087), HIF1A antibody (no./AF1009), claudin-5
(no./AF5216), and VEGFA antibody (no.: DF7470)—Affinity, China;
Alexa Fluor 555-labeled donkey anti-mouse IgG(H + L) (A0460), Triton
X-100 (no.: ST795), and antifade mounting medium (no.: P0126)—Beyotime
Biotechnology, China; neutral balsam (no.: G8590, Beijing Solarbio
Science & Technology Co., Ltd. China); RNA Easy Fast Tissue/Cell
Kit (no.: DP451), FastKing gDNA Dispelling RT SuperMix (no.: KR118),
and Talent qPCR PreMix (SYBR Green) (no.: FP209)—Tiangen Biotech
(Beijing) Co., Ltd, China; anti-GAPDH (no: AF7021, Affinity); and
BCA Protein Assay Kit (no: P0012S, Beyotime Biotechnology, China).
3
Assessing Inflammatory Responses in Cell Cultures
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Azocasein was purchased from Sigma Chemical Co. (St. Louis, MO, USA). 4-nitro-3-octanoyloxybenzoic acid (NOB) was purchased from Abcam (Shanghai, China). Protocatechuic acid (PCA), Gentisic acid (DHB), the BCA Protein Assay Kit, the Mouse IL-6 or TNF-α ELISA KIT, and the Human IL-6 or TNF-α ELISA KIT were purchased from Solarbio Science & Technology Co., Ltd. (Beijing, China). Sparkjade ECL super and Color enhanced Prestained Protein Ladder (10–180 kD) were purchased from Sparkjade Biotechnology Co., Ltd. (Jinan, China). NF-κB p65 (D14E12) XP® Rabbit mAb (#8242), p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb (#4695), IκBα (44D4) Rabbit mAb (#4812), p38 MAPK (D13E1) XP® Rabbit mAb (#8690), β-actin (13E5) Rabbit mAb (#4970), and Anti-rabbit IgG HRP-linked Antibody (#7074) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). VEGFA Antibody (#AF5131) and FGF2 Antibody (#DF6038) were purchased from Affinity Biosciences (Changzhou, China). HaCaT cells were obtained from China Infrastructure of Cell Line Resource (Beijing, China). All other regents used were of analytical grade.
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