We used antibodies to FABP4 (Abcam, ab92501, WB 1:1000 dilution; IF 1:50 dilution), fibronectin (Abcam, ab45688, 1:5000 dilution), α-SMA (Abcam, ab124964, 1:10,000 dilution), collagen-I (Proteintech, 14695-1-AP, 1:1000 dilution), TGF-β1 (Proteintech, 21898-1-AP, 1:1000 dilution), p-Smad2 (CST, 3104 S, 1:1000 dilution), p-Smad3 (CST, 9520 T, 1:1000 dilution), PGC1α (Abcam, ab54481, 1:1000 dilution), PPARγ (Proteintech, 16643-1-AP, 1:1000 dilution), caspase-3 (Proteintech, 19677-1-AP, 1:1000 dilution), Bcl-2 (Santa Cruz, sc-7382, 1:1000 dilution), BIP (CST, 3177 S, 1:1000 dilution), CHOP (CST, 2895 T, 1:1000 dilution). Recombinant human TGF-β1 (Peprotech, 100-21C-10), Oil Red O (Sigma-Aldrich, O0625). FABP4 inhibitor BMS309403 (R&D systems, 5258/10).
Ab45688
Manufactured by Abcam
Sourced in United Kingdom, United States
Ab45688 is a monoclonal antibody that targets a specific protein. It is designed for use in various laboratory applications, such as immunoassays and immunohistochemistry. The product details and technical specifications are available on the Abcam website.
Automatically generated - may contain errors
Lab products found in correlation
Ab34710, by Abcam (9 mentions)
Ab124964, by Abcam (6 mentions)
Ab6586, by Abcam (6 mentions)
Ab205718, by Abcam (5 mentions)
Ab7817, by Abcam (4 mentions)
Ripa lysis buffer, by Beyotime (4 mentions)
Ab124964, by Proteintech (3 mentions)
E cadherin, by Cell Signaling Technology (3 mentions)
Ab260043, by Abcam (3 mentions)
Ab32575, by Abcam (3 mentions)
Ab8227, by Abcam (3 mentions)
Ab125066, by Abcam (3 mentions)
Chemidoc xrs imaging system, by Bio-Rad (3 mentions)
Ab54481, by Proteintech (2 mentions)
Oil red o, by Merck Group (2 mentions)
Ab216347, by Abcam (2 mentions)
β actin, by Cell Signaling Technology (2 mentions)
E cadherin, by BD (2 mentions)
Ab18197, by Abcam (2 mentions)
Ab1416, by Abcam (2 mentions)
51 protocols using ab45688
1
Adipogenesis Regulation and Signaling
2
Multiplex Immunofluorescence Profiling of Lymph Nodes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fresh LNs were embedded in tissue-freezing medium. Cryostat sections (8 μm thick) were cut for imaging by fluorescence confocal microscopy. The following primary Abs were used for tissue staining: anti-HEV MECA79 (sc-19602, SCBT), anti-Lyve-1 (ab14917, Abcam), anti-PDPN (AF3244, R&D Systems), anti-ER-TR7 (sc-73355, SCBT), anti-CD11b (101202, BioLegend), anti-PDGFRβ (136005, Biolegend), anti-NG2 (ab129051, Abcam), anti-RANKL (510002 Biolegend), anti-CD35 (NBP2-52667, Novus Biologicals), anti-MAdCAM (16-5997-85,eBioscience), anti-ZO-1 (61-7300, Invitrogen), anti-Collagen I (ab34710, Abcam), anti-Fibronectin (ab45688, Abcam), anti-LepR (L9536, Sigma-Aldrich). The following secondary Abs were used: Alexa Fluor 488-conjugated anti-rabbit IgG, Alexa Fluor 594-conjugated anti-rabbit IgG, Alexa Fluor 488-conjugated anti-rat IgG, Alexa Fluor 594-conjugated anti-rat IgG, Alexa Fluor 488-conjugated anti-goat IgG, and Alexa Fluor 594-conjugated anti-goat IgG (Jackson ImmunoResearch). The stained tissue sections were imaged using EVOS FL Auto 2 Imaging System (Thermo Fisher Scientific). For the quantification of images, all images were automatically processed using ImageJ (NIH) and split into RGB channels. Auto threshold was used to convert intensity values of the immunofluorescent stain into numeric data. DAPI (VECTASHIELD, Vector Laboratories) was used to stain the cell nuclei.
3
FN1 Expression in Bladder Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemical (IHC) staining for streptavidin-peroxidase (SP) was used to detect FN1 expression in bladder tissue. After sectioning, conventional dewaxing, gradient ethanol hydration, and antigen retrieval were performed in sequence. Subsequently, the sections were incubated with 3% hydrogen peroxide for 20 min, blocked with serum for 10 min, and incubated with the primary antibody rabbit anti-mouse FN1 (ab45688; Abcam, USA; 1:200) overnight at 4 °C. The sections were then incubated with horseradish peroxidase-labeled goat anti-rabbit secondary antibody (ab205718; Abcam, USA; 1:2000) for 30 min and stained with DAB (Sigma, USA). After counterstaining with hematoxylin, all slides were hydrated, clarified, fixed and observed, and the positive cell index was determined.
4
Quantitative Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Harvested cells or collected tissues were lysed in radioimmunoprecipitation assay (RIPA) buffer (Beyotime Biotechnology, Shanghai, China) with protease inhibitor (Sigma-Aldrich, Darmstadt, Germany). The protein was quantified using the bicinchoninic acid (BCA) protein assay (Thermo Scientific, Waltham, MA, USA). Total proteins were resolved by 12% Tris-glycine sodium dodecyl sulfate polyacrylamide gel electrophoresis and were transferred to a polyvinylidene fluoride membrane (Millipore, Darmstadt, Germany). Membranes were blocked with 5% nonfat dry milk in PBST for 1 h and then incubated with primary antibodies against mouse Fibronectin (ab45688, Abcam), TLR4 (ab13867, Abcam), NF-κB (#8242, CST), phospho-NF-κB p65 (#3031, CST), MyD88 (#4283, CST) or GAPDH (#5174, CST) followed by incubation with a peroxidase-conjugated goat anti-rabbit (or mouse) IgG antibody. Densitometry analysis was performed, and the results were normalized to GAPDH expression and expressed as the fold changes over controls.
5
Antibody Characterization and NFkB Inhibition
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Twist2 antibody was purchased from Abcam (#ab66031) and used at 1:5000 for immunoblotting, 1:1000 for immunohistochemistry and 1:500 for immunofluorescence. Antibodies for NFkB (1:1000; #4764), pNFkB (1:1000; #3033), ITGA1 (2 μg/ml; #71,747), Vimentin (1:5000; #9855), and E-cadherin (1:1000; #3195) were purchased from Cell Signaling Technology. The LAMA4 (1:1000; ab205568) and Fibronectin (1:15,000; ab45688) antibodies were purchased from Abcam. The RhoC antibody, used at a 1:1000 dilution, was graciously shared by Dr. Kenneth van Golen (University of Delaware). The β-actin antibody (1:10,000; #A5316) was used as a loading control for immunoblotting and was purchased from Sigma-Aldrich Corp. AlexaFluor488 anti-rabbit or AlexaFluor 568 anti-mouse secondary antibodies were used for Western blots scanned with the Typhoon FLA 9000. An AlexaFluor488 anti-rabbit secondary antibody was used for immunofluorescence. Inhibition of NFkB was performed using NFkB Activation Inhibitor II, JSH-23 (#481,408; Millipore) at a concentration of 10 μM for 2 h.
6
Western Blot Analysis of EMT Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The primary antibodies, including anti-Snail1 (ab216347, Abcam, Cambridge, MA, USA), anti-Zeb1 (ab203829, Abcam), anti-E-cadherin (ab40772, Abcam), anti-fibronectin (ab45688, Abcam), and anti-actin (ab179467, Abcam), were used for Western blot analysis. The Western blot analysis was performed as previously described [5 (link), 6 (link), 17 (link), 18 (link)].
7
Western Blot Analysis of Notch Pathway Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analysis was performed as described previously [25 (link)]. In brief, protein lysates of cultured cells were prepared using tissue protein extraction reagent (Pierce, Rockford, IL, USA) according to the manufacture’s instruction. Equal amounts of protein lysates were subjected to SDS-polyacrylamide gel electrophoresis. After gel electrophoresis, the separated proteins were transferred onto a polyvinylidene difluoride (PVDF) membrane (Bio-Rad) and were then probed with specific primary antibodies. Primary antibodies against fibronectin (ab45688; Abcam; 1:2000 dilution), α-SMA (ab7817; Abcam; 1:2000 dilution), E-cadherin (#610181; BD Biosciences; 1:2000 dilution), NICD2 (#5732S; Cell Signaling; 1:1000 dilution), NICD1 (#2421S; Cell Signaling; 1:750 dilution), nicastrin (ab68145; Abcam; 1:1000 dilution), MAML-1 (ab65090; Abcam; 1:1000 dilution), Myc-tag (ab9106; Abcam; 1:2000 dilution), Jagged-1 (sc-390177; Santa Cruz; 1:1000 dilution), Delta-like 1 (#2588; Cell Signaling; 1:1000) and β-actin (#4970S; Cell Signaling; 1:6000 dilution) were obtained commercially. Secondary antibodies used for protein detection included goat anti-rabbit IgG-Horseradish Peroxidase (sc-2030; Santa Cruz; 1:3000 dilution) or goat anti-mouse IgG-HRP (sc-2005; Santa Cruz; 1:3000 dilution).
8
Western Blot Analysis of Fibrosis Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The expression levels of α-SMA, COL1α1, FN1, and vimentin were examined by WB. The following antibodies were used: anti-α-SMA (Abcam, ab124964), anti-COL1α1 (Abcam, ab260043), anti-FN1 (Abcam, ab45688), anti-vimentin (Abcam, ab92547) and anti-β-actin (Immunoway, YM3028). Details are provided in the Additional file 2 : Methods.
9
Glioma Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The proteins of glioma cells and tissues were extracted using radio-immunoprecipitation assay (Sigma-Aldrich) and denatured at 100°C before they were separated using sodium dodecyl sulphate-polyacrylamide gel electrophoresis. After transferring to the polyvinylidene fluoride (Sigma-Aldrich) membranes and immersing in 5% skimmed milk, the membranes were conjugated with primary antibodies against Wnt7B (ab94915, 1:1,000, Abcam, Cambridge, MA, USA), Total-caspase 3 (ab32351, 1:5,000), cleaved-caspase 3 (ab32042, 1:5,000), N-cadherin (ab76057, 1:1,500), E-cadherin (ab1416, 1:100), fibronectin (FN, ab45688, 1:1,000), and snail (ab180714, 1:1,000), proliferating cell nuclear antigen (PCNA) (ab18197, 1:2,000), or GAPDH (ab8245, 1:2,000) at 4°C overnight. Following, the membranes were mixed with corresponding secondary antibodies (ab6721, 1:20,000; ab6789, 1:15,000) for 1 h. Finally, an enhanced chemiluminescence Western blotting system (Bio-Rad, Hercules, CA, USA) was used to detect these membranes.
10
Immunohistochemical Analysis of Tissue Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Deparaffinized tissues were heated to expose the antigens using Tris-ethylene diamine tetraacetic acid (EDTA) retrieval solution (pH 6.0) at 95°C for 30 minutes. Wash with PBS for three times per 3 minutes and incubate the sections in 3% hydrogen peroxide (H2O2) to block endogenous peroxidase activity. And then, samples were stained using primary antibodies to detect α-SMA (Abcam, ab32575), FAP (Abcam, ab28244), vimentin (Abcam, ab92547), fibronectin (Abcam, ab45688), E-cadherin (Abcam, ab76319), VCAM-1 (Abcam, ab134047), collagen I (Abcam, ab138492), cytokeratin (Abcam, ab6401). Antibodies were followed by means of peroxidase-conjugated secondary antibody and revealed with 3-39-diaminobenzidine (DAB) as chromogen.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!