5-μm-thick paraffin sections were used for immunohistochemistry. Following deparaffinized, antigen repair and sealed off, the sections were incubated with antibody against PHGDH (1 : 200, 14719-1-AP, Proteintech) or Ki67 (1 : 200, 9027 T, CST) at 4 °C overnight. The slides were incubated with biotinylated goat anti-rabbit antibodies for 1.5 h, stained with diaminobenzidine (abs957, Absin Biotechnology Co., Ltd, Beijing, China) and then counterstained with hematoxylin (abs957, Absin Biotechnology Co., Ltd, Beijing, China). The sections were scored according to the percentage of positive staining cells (0 = negative; 1 = 5–25%; 2 = 26–50%; 3 = 51–74%; and 4 = 75–100%) and the intensity of staining (0 = no staining; 1 = slight staining; 2 = moderate staining; and 3 = strong staining). Scores for the percentage and intensity of staining were added.
Abs957
Manufactured by Absin
Sourced in China
The Abs957 is a laboratory equipment designed for general sample processing and analysis. It features a compact and durable construction, and is capable of performing essential tasks such as mixing, stirring, and controlled heating or cooling of samples. The Abs957 is a versatile tool suitable for use in various scientific and research settings.
Automatically generated - may contain errors
Lab products found in correlation
Vt1200, by Leica (1 mentions)
Anti iba1, by Fujifilm (1 mentions)
Anti gfap, by Merck Group (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
Hm 325, by Thermo Fisher Scientific (1 mentions)
Ab40759, by Abcam (1 mentions)
Fluorescence microscopy, by Olympus (1 mentions)
Cleaved caspase 3, by Absin (1 mentions)
6 protocols using abs957
1
Immunohistochemical Quantification of PHGDH and Ki67
2
Immunohistochemical and Ultrastructural Analysis of Gliosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After transcardial perfusion of buffered 4% PFA, brain tissues were immediately immersed in 0.1 M PB containing 4% PFA and 0.5% glutaraldehyde (pH 7.4) at 4 °C for 6 h, and 200-µm-thick slices were cut with a vibratome (VT-1200S, Leica) and then stained exactly as described in the DAB kit instructions (abs957, Absin, Shanghai, China). The brain slices were incubated with anti-Iba-1 (1:200, WAKO, Tokyo, Japan) and anti-GFAP (1:200, Millipore) primary antibody and secondary antibody conjugated with HRP. Immunoreaction products were visualized using DAB substrate. Following observation under a light microscope and identification of the gliosis region, tissue pieces were cut into 1 mm × 1 mm, additionally fixed with 2.5% glutaraldehyde, and prepared for TEM imaging. In brief, tissues were washed with PB, treated with 1% OsO4 for 90 min at RT. Then tissues were dehydrated in a graded series of ethanol (50%, 70%, 90%), and incubated with 100% acetone, a 1:1 mixture of epoxy resin and acetone, and 100% resin (overnight). Tissues were embedded in epoxy resin and cured in a 60 °C oven for 72 hrs. In all, 1-µm-thick sections were cut and stained with 0.1% toluidine blue to distinguish the target region with a light microscope. 100 nm-ultrathin sections were stained with lead citrate and examined under TEM.
3
Immunohistochemical Analysis of Angiogenesis
4
Histological Analysis of Ly6G+ Cells in Mouse Maxillae
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples were prepared from maxillae for histological analyses. After fixing the samples with 4% paraformaldehyde for 24 hours, decalcification solution (Yobibio, YB2003) was used for demineralization at 4°C, and the solution was changed every two days. Paraffin embedding was carried out one month later. Paraffin-embedded blocks were sectioned at 5μm intervals using a microtome (HM325, Thermo Fisher Scientific, USA). Ly6G+ cells in the mouse maxillae were examined by immunohistochemistry (IHC) with a the specific primary antibody (1:500; Abcam, ab238132) and a diaminobezidin (DAB) system (Absin, abs957) according to the manufacture’s instruction. Five high-power fields (hpf) (×400) per each sample at ROI were randomly selected and the positively-stained cells were enumerated by Image Pro Plus 6.0 (Media Cybernetics, Silver Spring, MD).
5
Immunohistochemical Analysis of TGF-β Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The sections were stained by IHC to detect the expression of factors in the TGF-β family. After the antigen was repaired, primary antibodies were added at room temperature for 20 min and then secondary antibodies for 10 min, avoiding light. Diaminobenzidine (DAB) was used for staining, and neutral gum was used to seal pieces. The pieces were observed under light microscope, and 3 fields were collected from each section. ImagePro Plus software was adopted to calculate the average optical density. The antibodies were TGF-Beta1 antibody (Proteintech, 21898-1-AP), anti-TAK1 (Rabbit Monoclonal) (Abcam, ab109526), JNK antibody (Rabbit Polyclonal) (Proteintech, 10023-1-AP), FSHR antibody(Proteintech, 22665-1-AP), Anti-Smad4 (Abcam, ab40759), and a high-efficiency immunohistochemical secondary antibody kit (Absin, abs957).
6
Renal Tissue Immunohistochemistry Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Renal samples were collected and fixed in 4% paraformaldehyde at room temperature for 24 h, paraffin embedded, and sectioned at 4 µm. Then, the slides were deparaffinized, rehydrated in a descending alcohol series and the antigen was exposed by 20 min of incubation at 100°C in sodium citrate. Thereafter, the slides were incubated with 3% H2O2 at 37°C for 10 min to block the endogenous peroxidase activity. Sequentially, the slides were incubated with bovine serum albumin (cat. no. abs957; Absin Bioscience Inc., Shanghai, China) and the primary antibodies (cleaved Caspase 3, cat. no. 9664, 1:200, Cell Signaling Technology, Inc., Danvers, MA, USA; MPO, cat. no. abs120616, 1:200, Absin Bioscience Inc.) overnight at 4°C. After incubation with biotin-labeled secondary antibody (abs957, Absin Bioscience Inc.) at room temperature for 10 min, the color was developed with 3,3′-diaminobenzidine at room temperature for 30 sec. Then, the tissues were imaged by fluorescence microscopy (Olympus Corporation). The staining was evaluated in a blinded manner.
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!