Ab116222

Manufactured by Abcam

Sourced in United Kingdom

Ab116222 is a laboratory reagent designed for use in research applications. It is a monoclonal antibody that targets a specific antigen. The core function of this product is to serve as a tool for researchers to detect and study the targeted antigen in their experiments.

Automatically generated - may contain errors

Lab products found in correlation

Ab119716, by Abcam (1 mentions) Ab6302, by Abcam (1 mentions) Ab129119, by Abcam (1 mentions) Sds page, by Solarbio (1 mentions) Cw2334s, by CWBIO (1 mentions) Cw2200, by CWBIO (1 mentions) Ipvh00010, by Merck Group (1 mentions) Cw0014, by CWBIO (1 mentions) Fluorescent secondary antibody, by LI COR (1 mentions) Anti phospho gsk3β, by Cell Signaling Technology (1 mentions) Anti β catenin, by Cell Signaling Technology (1 mentions) Anti gsk3β, by Cell Signaling Technology (1 mentions) Anti phospho β catenin, by Cell Signaling Technology (1 mentions) Bca assay kit, by Thermo Fisher Scientific (1 mentions) Ab8227, by Abcam (1 mentions) Anti β actin, by Abcam (1 mentions) Ab260043, by Abcam (1 mentions) Wbkls0100, by Merck Group (1 mentions) P0013, by Beyotime (1 mentions) P0012, by Beyotime (1 mentions)

4 protocols using ab116222

Immunohistochemical Evaluation of WNT3 in Placenta

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunohistochemical staining was used to evaluate the location and expression of WNT3 proteins in placental tissues. Placental tissue sections were heated for 2 h at 60 °C, deparaffinized by immersion in xylene two times for 10 min each, and dehydrated through a series of graded ethanol solutions (100, 95, 85 and 75%).
Sections were immersed in the prepared antigen retrieval solution and were then sequentially heated to greater than 90 °C in a microwave oven and then quenched at a low temperature for 20 min. Furthermore, the sections were incubated with an anti-Wnt3 antibody (1:200; ab116222; Abcam) at 4 °C overnight. After washing with PBS three times, the sections were incubated with a biotin-conjugated secondary antibody (1:200; OriGene Technologies, Inc.) for 1 h at room temperature. The sections were stained with DAB reagent, counterstained with hematoxylin, and finally sealed with neutral balsam. The slides were examined by inverted fluorescence microscopy (OLYMPUS IX-71, Tokyo, Japan). The staining of the sections was independently evaluated by two pathologists.

Zhang L., Sang M., Li Y., Li Y., Yuan E., Yang L., Shi W., Yuan Y., Yang B., Yang P, & Yuan E. (2021). WNT3 hypomethylation counteracts low activity of the Wnt signaling pathway in the placenta of preeclampsia. Cellular and Molecular Life Sciences, 78(21-22), 6995-7008.

+ Open protocol

+ Expand

Analyzing Intestinal Wnt Signaling Pathway

Check if the same lab product or an alternative is used in the 5 most similar protocols

For the extraction of total protein, 100 μL RIPA lysis buffer (CW2334S; CWBIO, Beijing, China) and 1 μL Protease Inhibitor Cocktail (CW2200S; CWBIO, Beijing, China) were added to 100 mg of jejunal tissue, and the total protein was quantified using a bicinchoninic acid protein assay kit (CW0014; CWBIO, Beijing, China). Protein was separated by SDS-PAGE (Solarbio, Beijing, China) (20 μg per lane) and then transferred onto PVDF membranes (IPVH00010; Millipore, Danvers, MA). Next, the membranes were incubated with polyclonal goat anti-Wnt3 (ab116222; Abcam, UK), polyclonal rabbit anti-β-catenin (ab6302; Abcam, UK), monoclonal rabbit anti-TCF4/TCF7L2 (2565; CST, Boston, MA), polyclonal rabbit anti-ZO-1 (61-7300; Invitrogen, Camarillo, CA), and polyclonal rabbit anti-Claudin (ab129119; Abcam, UK) (all 1:2000 in Tris-buffered saline with Tween) overnight at 4°C. The membranes were incubated with HRP-conjugated rabbit anti-goat IgG (H&L) (1:5000; bs-0294R; Beijing Bioss Biotechnology Co., Ltd., Beijing, China) or HRP-conjugated goat anti-rabbit IgG (H&L) (1:5000; K008; Kmbio, Beijing, China) for 1 h at 37°C. Polyclonal anti-β-actin (1:5000; ab119716; Abcam, UK) was used for normalization of band intensities. The densitometric values of obtained immunoblot signals from 6 separate experiments were determined using Image J (National Institutes of Health, New York).

Deng Z., Han D., Wang Y., Wang Q., Yan X., Wang S., Liu X., Song W, & Ma Y. (2021). Lactobacillus casei protects intestinal mucosa from damage in chicks caused by Salmonella pullorum via regulating immunity and the Wnt signaling pathway and maintaining the abundance of gut microbiota. Poultry Science, 100(8), 101283.

+ Open protocol

+ Expand

Protein Extraction and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

RIPA lysis buffer containing a protease inhibitor (CWBIO, China) was used to extract protein from placental tissues and cells. The total protein concentration was measured with a BCA assay kit (Thermo Fisher Scientific, Inc.). For western blot analysis, approximately 40 μg of protein was separated on an 8% SDS-PAGE gel and electrophoretically transferred to PVDF membranes. The membranes were blocked for 2 h at room temperature with blocking buffer (5% nonfat milk, 0.1% Tween 20). Then, the membranes were incubated at 4 °C overnight with anti-Wnt3 (1:1000; ab116222; Abcam), anti-β-catenin (1:1000; Cell Signaling Technology, Inc.), anti-phospho-β-catenin (1:1000; Cell Signaling Technology, Inc.), anti-GSK3β (1:1000; Cell Signaling Technology, Inc.), anti-phospho-GSK3β (1:1000; Cell Signaling Technology, Inc.), and anti-β-actin (1:2000; ab8227; Abcam) antibodies separately. The membranes were then incubated with fluorescent secondary antibodies (1:15,000; LI-COR, USA) for 2 h at room temperature. An infrared laser scanning imaging system (Odyssey CIX, LI-COR, USA) was used to determine the fluorescence intensity.

+ Open protocol

+ Expand

Protein Expression Analysis in MC3T3-E1 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Following the designated pharmacological treatments, MC3T3-E1 cells were lysed and harvested in the WB and IP cell lysis buffer (P0013, Beyotime, Shanghai, China) for protein extraction. Subsequently, the bicinchoninic acid (BCA) quantification assay (P0012, Beyotime) was employed to estimate the protein concentrations. Following that, the total protein, amounting to 20 μg, underwent SDS-PAGE for analysis. The resultant protein bands were transferred onto a PVDF membrane. In an ambient environment, the membrane was blocked using 5% skimmed milk for 1 h. Thereafter, the diluted primary antibodies, including anti-collagen I (diluted at 1:1000, ab260043, Abcam), RUNX2 (diluted at 1:1000, 12556, CST), anti-Wnt3 antibody (diluted at 1:1000, ab116222, Abcam), β-catenin (diluted at 1:1000, 8480, CST), and β-actin (diluted at 1:2000, 3700, CST; loading control) were added and incubated overnight at 4 °C. The PVDF membrane was subjected to three washes with 1 × TBST. Subsequently, another incubation at ambient temperature lasting 1 h was carried out employing horseradish peroxidase-conjugated secondary antibodies targeting either rabbit or mouse (7074, 7076, CST). Finally, the membrane and ECL (WBKLS0100, Merck, Germany) were incubated, and the chemiluminescent spectra were captured and illuminated using the imaging system (Bio-Rad, Hercules, CA, USA).

Zhao J., Liang G., Yang J., Huang H., Dou Y., Gu Z., Liu J., Zeng L, & Yang W. (2024). Liuwei Dihuang Pills Enhance Osteogenic Differentiation in MC3T3-E1 Cells through the Activation of the Wnt/β-Catenin Signaling Pathway. Pharmaceuticals, 17(1), 99.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!