To detect endogenous IL23A secretion and binding region in brain, we performed AP-binding analysis. Briefly, we obtained 10-μm fresh frozen coronal sections and postfixed with precooled methanol at −20°C for 8 min. The sections were incubated with biotinylated IL23A antibody (Abcam, ab119545) overnight at 4°C and then washed with HBAH buffer (0.5 mg/mL BSA, 0.1% [w:v] NaN3, 20 mmol/L HEPES in Hanks’ balanced salt solution, pH 7.0). After incubating with streptavidin (SA)-AP (Invitrogen, 434,322) for 1.5 h at room temperature, the sections were fixed with 4% PFA for 5 min and washed with HBS buffer (150 mmol/L NaCl, 20 mmol/L HEPES, pH 7.0). Finally, inactivating endogenous AP activity at 65°C for 1 h and AP binding was detected by BCIP/NBT substrate solution (Sangon Biotech, E670032) according to the manufacturer.
Ab119545
Manufactured by Abcam
Sourced in United Kingdom
Ab119545 is a recombinant monoclonal antibody that recognizes human EGFR. The antibody is produced in HEK293 cells.
Automatically generated - may contain errors
3 protocols using ab119545
1
Mapping Endogenous IL23A in the Brain
2
Evaluating Anti-Inflammatory Effects of HGs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess the anti-inflammatory effect of 10HA and 10HA 120NP HGs further, ELISA assays were performed to quantify the release of six pro-inflammatory mediators by LPS-stimulated RAW264.7. Mouse TNF-α, IL-6, IL-10, IL-12p40, IL-23, and PGE2 ELISA kits were purchased from abcam (ab208348, ab222503, ab255729, ab236717, ab119545, and ab133021, Cambridge UK). In brief, cells were seeded on top of the HGs and CGs at a density of 2 × 106 cells/well and cultured for 24 h. Then, cells were exposed to medium containing LPS (500 ng/mL) and incubated overnight. Media were collected and stored at −20 °C until use. Levels of mouse TNF-α, IL-6, IL-10, IL-12p40, IL-23, and PGE2 in cell culture supernatants were determined by the corresponding ELISA kit according to the protocol recommended by the manufacturer. Experiments were performed using six replicates per formulation. ANOVA was performed at significance levels of p < 0.05, p < 0.01, and p < 0.001.
3
Serum Cytokine Quantification by ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Enzyme-linked immunosorbent assay was used to determine serum concentrations of the IL-23 (ab119545, Abcam, UK) and IL-17 by ELISA kits (ab100702, Abcam, UK) according to the manufacturers' instructions. The optimal density at 450 nm was used to measure the concentration of IL-23 and IL-17 in a microplate reader. The curve equation and r value were calculated to determine the concentrations of samples.
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!