The harvested subcutaneous tumors were fixed with 4% paraformaldehyde (Solarbio, Beijing, China) and embedded in paraffin, finally sliced into 6 µm-thick sections for IHC analysis. Briefly, these tissues sections were deparaffinized in xylene for 30mins and hydrated in a descending alcohol series (100% alcohol, 95% alcohol, 80% alcohol) for 5 min respectively. After antigen retrieval with EDTA (ZSGB-BIO, China) and blocking with goat serum for one hour at room temperature, the sections were incubated with the corresponding antibody at 4 °C overnight. Followed by incubation with enhanced enzyme-labeled goat anti-mouse/rabbit IgG polymer (ZSGB-BIO, China), IHC signal was detected by 3,3′-diaminobenzidine (DAB, ZSGB-BIO, China). The antibodies used above were: anti-CD4 (1:500, ab183685, Abcam), anti-CD8 (1:500, ab217344, Abcam), anti-CD163(1:500, ab182422, Abcam), anti-E-cadherin (1:500, 3195T, Cell Signaling Technology), anti-IFN-γ (1:100, AF-585-NA, R&D), anti-DCLK1 (1:500, ab31704, Abcam)
Af 585 na
Manufactured by R&D Systems
Sourced in United States
AF-585-NA is a laboratory instrument designed to measure the concentration of specific analytes in a sample. It utilizes a proprietary detection method to provide accurate and reliable results. The core function of this product is to enable researchers and scientists to quantify the presence of target molecules within their experimental samples.
Automatically generated - may contain errors
Lab products found in correlation
Ab217344, by Abcam (1 mentions)
Ab31704, by Abcam (1 mentions)
Ab182422, by Abcam (1 mentions)
Ab183685, by Abcam (1 mentions)
Ethylene diamine tetraacetic acid (edta), by ZSGB-BIO (1 mentions)
Paraformaldehyde (pfa), by Solarbio (1 mentions)
3 3 diaminobenzidine dab, by ZSGB-BIO (1 mentions)
Brain derived neurotrophic factor (bdnf), by Merck Group (1 mentions)
Sp600125, by Apexbio (1 mentions)
Bay11 7082, by Apexbio (1 mentions)
Pd98059, by Apexbio (1 mentions)
Y 27632, by Apexbio (1 mentions)
Ag490, by Apexbio (1 mentions)
Te300, by Nikon (1 mentions)
U0126, by Apexbio (1 mentions)
Ly294002, by Apexbio (1 mentions)
2 protocols using af 585 na
1
Immunohistochemical Analysis of Subcutaneous Tumors
2
Modulation of Cell Signaling Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PC12 or C17.2 cells were seeded in a 6-well plate at the density of 1 × 106 cells/well and incubated for 48 h until reaching 80 % confluence. Then, cells were treated with 10% CM (v/v) from M. smegmatis, or 0.5 ng/ml interferon gamma (IFN-γ) (ab645, abcam, USA), or anti-INFγ (1:1,000 dilution, AF-585-NA, R&D system, USA) for 48 h. For positive control, PC12 cells were treated with 50 ng/ml nerve growth factor (NGF Sigma, USA), and C17.2 cells were treated with 50 ng/ml NGF and 50 ng/ml brain-derived neurotrophic factor (BDNF, Sigma, USA. For screening experiment of signal pathway, cells were treated with CM and different inhibitors (AG490, SP600125, U0126, SB239063, PD98059, H89, Y27632, IWP2, DAPT, BAY 11-7082, LY294002, APExBIO Technology, Houston, USA) for 48 h respectively. The morphology of the cells was observed by microscope (Nikon TE 300).
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!