Liver tissues were fixed overnight in 10% formaldehyde and were embedded in paraffin. Slides were stained with H&E, Masson's Trichrome, or Picric acid-Sirius red. For immunohistochemical analysis, paraffin-embedded or frozen sections were incubated with antibodies against mouse CD45 (30-F11; BD Bioscience), CD68 (KP1; Abcam), MPO (Rabbit polyclonal; Abcam), Dectin-1 (R1-8g7; Invivogen), PCNA (PC10; Biolegend), TLR4 (Rabbit polyclonal; Abcam), α-SMA (1A4; Abcam), Phalloidin (Cell Signaling), or M-CSF (Rabbit polyclonal; Abcam). Human liver sections were stained with an antibody against Dectin-1 (Rabbit polyclonal; Abcam) or CD11b (M1/70; Biolegend). Quantification was performed by examining 10 high powered fields (HPFs) per slide. Fibrosis was quantified based on Trichrome staining using a computerized grid as described (Ochi et al., 2012a (link)). Immunofluorescent imaging was performed using a LSM 700 confocal microscope and an Axiovert camera (Zeiss).
Dectin 1
Manufactured by Abcam
Sourced in United Kingdom, China
Dectin-1 is a type II transmembrane receptor expressed on the surface of myeloid cells, such as macrophages and dendritic cells. It functions as a pattern recognition receptor that binds to beta-glucans, a component of fungal cell walls, and plays a role in the innate immune response to fungal infections.
Automatically generated - may contain errors
Lab products found in correlation
P syk, by Cell Signaling Technology (2 mentions)
β actin 8h10d10, by Cell Signaling Technology (2 mentions)
P plcγ, by Cell Signaling Technology (2 mentions)
Rb c 15, by Cell Signaling Technology (2 mentions)
Galectin 9, by InvivoGen (2 mentions)
Dectin 1 fc fc mdec1a, by InvivoGen (2 mentions)
Bcl xl 54h6, by Cell Signaling Technology (2 mentions)
C myc, by Cell Signaling Technology (2 mentions)
Rneasy mini kit, by Qiagen (2 mentions)
Dc protein assay, by Bio-Rad (2 mentions)
Ncounter analysis system, by NanoString (2 mentions)
Lsm 700 confocal microscope, by Zeiss (2 mentions)
Phalloidin, by Cell Signaling Technology (1 mentions)
Cd45 30 f11, by BD (1 mentions)
Cd68 kp1, by Abcam (1 mentions)
α sma 1a4, by Abcam (1 mentions)
Cd11b m1 70, by BioLegend (1 mentions)
Axiovert camera, by Zeiss (1 mentions)
M csf, by Abcam (1 mentions)
β tubulin, by Elabscience (1 mentions)
6 protocols using dectin 1
1
Liver Tissue Histopathology and Immunohistochemistry
2
Protein Extraction and Western Blotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For protein extraction, tissues were homogenized in ice-cold RIPA buffer. Total protein was quantified using the DC Protein Assay according to the manufacturer’s instructions (BioRad, Hercules, CA). Western blotting was performed as previously described with minor modifications13 (link). Briefly, 10 % Bis-Tris polyacrylamide gels (NuPage, Invitrogen) were equiloaded with 10–30μg of protein, electrophoresed at 200V, and electrotransferred to PVDF membranes. After blocking with 5% BSA, membranes were probed with primary antibodies to β-actin (8H10D10), p53 (7F5), PLC-γ (polyclonal), p-PLC-γ (polyclonal), Bcl-XL (54H6; all Cell Signaling), JNK (2C6), p-JNK (G9), Smad4 (polyclonal), p16 (polyclonal), c-Myc (9E10), CARD9 (polyclonal), Syk (polyclonal), p-Syk (polyclonal), Rb (C-15; all Cell Signalling), Dectin-1 (polyclonal; Abcam), Galectin-9 (polyclonal), and Dectin-1 Fc (fc-mdec1a; InvivoGen). Blots were developed by ECL (Thermo Scientific, Asheville, NC). RNA extraction was performed using the RNeasy Mini kit (Qiagen, Germantown, MD) as per manufacturer’s instructions. For Nanostring analysis, the nCounter mouse inflammation panel was employed using the nCounter Analysis System (both Nanostring, Seattle, Washington).
3
Protein Extraction and Western Blotting
4
Investigating Corneal Protein Expression in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HCECs were observed under different inverted microscopes. Cells were seeded for adherent culture in 6-well plates, and allowed to reach approximately 80% confluence. The original culture solution was discarded. Three milliliters of pre-cooled PBS were added to each well to wash the cells. Then, cell lysis buffer was prepared with PMSF and 150 uL of this buffer was added to each well. Corneal tissues from B6 mice were obtained for Western blot assay. The membranes containing separated proteins were incubated with primary antibodies against Dectin-1 (1:1000), HO-1 (1:250), Nrf2 (1:1000), TNF-α (1:500; Abcam, Cambridge, UK), IL-1β (1:1000), IL-6 (1:1000), β-tubulin (1:1,000; Elabscience, Wuhan, China), and β-actin (1:1000) (Elabscience). Goat anti-rabbit (1:5000) and goat anti-mouse (1:800) secondary antibodies were also used. Next, the bands were detected with ECL Western blot detection reagents (Biotime, China) in accordance with standard protocols.
5
Western Blot Analysis of Immune and Apoptosis Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The brain tissues were cut, grinded, and lysed with RIPA lysis and extraction buffer (Cat #P0013B, Beyotime), and protease and phosphatase inhibitor were added (Cat #P105539, Aladdin). Proteins (30 μg) were separated by SDS-PAGE (5% concentrated gel and 12% separating gel) and transferred to PVDF membranes (Cat #ISEQ15150 and Cat #IPVH00010, Millipore). 5% nonfat milk was used to block, and the PVDF membranes were incubated with targeted primary antibodies and secondary antibodies. At last, the protein bands were visualized with an ECL detection solution (Thermo). The primary antibodies were as follows: rabbit polyclonal antibody dectin-1 (Cat #Ab140039, 1 : 1000; Abcam), mouse monoclonal antibody Bcl-2 (Cat #Ma1-12246, 1 : 1000, Thermo), mouse monoclonal antibody Bax (Cat #Sc-7480, 1 : 1000, Santa), rabbit polyclonal antibody caspase-3 (Cat #9662, 1 : 1000, Cst), and anti-GAPDH (Cat #AB-P-R 001, 1 : 5000, Hangzhou Xianzhi Biology Co., Ltd). We quantified the intensity of the protein bands with ImageJ software, while all the target proteins normalized to GAPDH.
6
Immunomodulatory Compound Screening Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BMS202, SC79, and NSC87877 were purchased from Selleck Chemicals (Houston, TX, USA); TNF-α, IL-6, and IL-1β enzyme-linked immunosorbent assay (ELISA) kits from Elabscience Biotechnology (Wuhan, China); PD-1 ELISA kits from Meimian (Jiangsu, China); HIF-1α, SHP1, SHP2, p-Akt, Akt, and β-actin from Cell Signaling Technology (Danvers, MA, USA); PD-1, CD11c, CD16, CD86, Anti-CD86/PE, CD206, Anti-CD206/FITC, and Arg-1 antibodies from Bioss (Beijing, China); Dectin-1 from Abcam (Cambridge, UK); uorophore-labeled goat anti-rabbit secondary antibody (Alexa Fluor 594) and goat anti-rabbit immunoglobulin horseradish peroxidase (IgG-HRP), BSA, and bicinchoninic acid assay (BCA) protein detection kits from Beyotime (Shanghai, China); SP (mouse/rabbit IgG)-POD kit from Solarbio Life Sciences (Beijing, China); FBS and Ham's F-12K (Kaighn's) medium from Gibco (Carlsbad, CA, USA).
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!