Ab170941

Manufactured by Abcam

Sourced in United Kingdom, United States

Ab170941 is a laboratory equipment product. It serves a core function, but a detailed description while maintaining an unbiased and factual approach is not available.

Automatically generated - may contain errors

Lab products found in correlation

Ab8805, by Abcam (5 mentions) Pvdf membrane, by Merck Group (4 mentions) Ab6721, by Abcam (3 mentions) Ripa lysis buffer, by Beyotime (3 mentions) Ab38449, by Abcam (3 mentions) P akt, by Cell Signaling Technology (3 mentions) Ab32503, by Abcam (3 mentions) Ab181602, by Abcam (2 mentions) Ab182651, by Abcam (2 mentions) Polyvinylidene fluoride membrane, by Merck Group (2 mentions) Ab2302, by Abcam (2 mentions) Ab134175, by Abcam (2 mentions) Ab8226, by Abcam (2 mentions) Ab81283, by Abcam (2 mentions) Ab8245, by Abcam (2 mentions) Ab34712, by Abcam (1 mentions) Alexa fluor 488, by Abcam (1 mentions) Confocal laser scanning microscope, by Olympus (1 mentions) Ab150077, by Abcam (1 mentions) Ab6785, by Abcam (1 mentions)

32 protocols using ab170941

Immunofluorescence Staining of RAW264.7 Macrophages

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

RAW264.7 cells were fixed with 4% paraformaldehyde for 20 min at room temperature. Then, cells were blocked with 5% goat serum for 1 h and incubated with primary antibodies against iNOS (1:80; ab170941; Abcam) and Arg‐1 (1:100; ab170941; Abcam) overnight at 4°C. On the next day, the membranes were cultivated with goat anti‐rabbit secondary antibody (1:100; ab6721; Abcam) for 1 h at room temperature. diamidino‐phenyl‐indole (DAPI) staining was utilized for cell dyeing for 5 min in the dark. The cells were photographed under a fluorescence microscope after blocking with anti‐fade mounting medium. The fluorescence intensity was observed using Image J software (version 1.46; National Institutes of Health).
For type II collagen, RAW264.7 cells were fixed with 4% paraformaldehyde for 20 min at room temperature. Then, cells that blocked with 5% goat serum for 1 h were incubated with primary antibody against Collagen II (1:100; ab34712 Abcam) overnight at 4°C, after which was the cultivation with a fluorescent goat anti‐rabbit IgG H&L (Alexa Fluor® 488) secondary antibody (1:100; ab150077 Abcam). DAPI staining was utilized for cell dyeing for 5 min in the dark. The fluorescence was observed using a laser scanning confocal microscope (magnification; x200; Olympus Corporation). The fluorescence intensity was observed using Image J software (version 1.46; National Institutes of Health).

Hou X., Shen Y., Sun M., Zhang B., Dai J., Chen D, & Liu Z. (2022). Effect of regulating macrophage polarization phenotype on intervertebral disc degeneration. Immunity, Inflammation and Disease, 10(11), e714.

+ Open protocol

+ Expand

PTEN Immunohistochemistry Staining Protocol

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

The paraffin-embedded sections were dehydrated by an ascending series of ethanol and washed with running water for 2 min. Then, the sections were immersed in 3% H₂O₂ for 20 min, followed by washing with distilled water for 2 min and 0.1 M PBS for 3 min. The samples were blocked by incubation in normal goat serum (C-0005; Shanghai Haoran Biological Technology, Shanghai, China) at room temperature for 20 min. Slides were incubated overnight at 4°C with the primary rabbit anti-human antibody to PTEN (1:100, ab170941; Abcam, Cambridge, UK). Following incubation with the secondary goat anti-rabbit antibody to immunoglobulin G (IgG; 1:1,000, ab6785; Abcam, Cambridge, UK) and a horseradish peroxidase (HRP)-labeled streptavidin working solution (0343-10000U; Imunbio, Beijing, China) at 37°C for 20 min, labeling was visualized using DAB (ST033; Whiga Biotech, Guangzhou, Guangdong, China) with hematoxylin (PT001; Bogoo Biotech, Shanghai, China). The sections were returned blue by adding 1% ammonia, then were dehydrated with gradient alcohol, cleared by xylene, and mounted with neutral resin. The section was observed and photographed under the optical microscope.

Li B., Luan S., Chen J., Zhou Y., Wang T., Li Z., Fu Y., Zhai A, & Bi C. (2019). The MSC-Derived Exosomal lncRNA H19 Promotes Wound Healing in Diabetic Foot Ulcers by Upregulating PTEN via MicroRNA-152-3p. Molecular Therapy. Nucleic Acids, 19, 814-826.

+ Open protocol

+ Expand

PTEN Immunohistochemistry in Nude Mice

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

Nude mice tumor tissues were fixed by paraformaldehyde solution, dewaxed in xylene, rinsed with phosphate buffered saline (PBS) and incubated with 3% hydrogen peroxide in 50% methanol for 30 minutes at 37°C. After the elimination of endogenous peroxidase activity, the sections were rinsed in PBS again and incubated in protein block solution (Bio-Genex, San Ramon, CA) in a humid chamber for 30 minutes. One hundred microliters PTEN was added to the sections, which were then incubated with primary antibody anti-ZFR (1:100, ab170941, Abcam) in a humid chamber overnight. Sections exposed to diluents alone without primary antibody served as negative controls. The slides were then rinsed three times in PBS for 5 minutes each and incubated with goat antimouse secondary antibody for 30 minutes. The reaction was developed using the peroxidase substrate diaminobenzidine. The sections were counterstained with hematoxylin.

Liu T., Liu S., Xu Y., Shu R., Wang F., Chen C., Zeng Y, & Luo H. (2018). Circular RNA-ZFR Inhibited Cell Proliferation and Promoted Apoptosis in Gastric Cancer by Sponging miR-130a/miR-107 and Modulating PTEN. Cancer Research and Treatment : Official Journal of Korean Cancer Association, 50(4), 1396-1417.

+ Open protocol

+ Expand

Protein Expression Analysis in Kidney Cells

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

Proteins from cultured cells or mice kidneys were extracted with TRI Reagent (Sigma–Aldrich) according to the manufacturer’s guidelines. Samples were separated by SDS-PAGE and transferred onto PVDF membranes (IPVH00010, Millipore). After blocking the membranes with 5% milk, we used primary antibodies to incubate the membranes overnight at 4 °C against the following proteins: KLF10 (1:1000, ab184182, Abcam), CCNB1 (1:1000, GTX100911, GeneTex), CCND1 (1:1000, GTX108624, GeneTex), PCNA (1:1000, 101239-T46, SinoBiological), KIM1 (1:1000, AF1817, R&D), Anti-FLAG (1:1000, 8146, CST), ACTIN (1:2000, GTX11003, GeneTex), ZBTB7A (1:1000, ab175918, Abcam), PTEN (1:1000, AB170941, Abcam), pAKT (1:1000, 4060 S, CST), AKT (1:1000, 4691 S, CST). The membranes were washed by TBST before incubated with secondary antibodies (1:2000, Jackson ImmunoResearch Inc). The bands of the target proteins were visualized by the LAS-3000 detection system and were quantitively analyzed by Fiji based on ACTIN and control groups respectively.

Zhang Y., Bao S., Wang D., Lu W., Xu S., Zhou W., Wang X., Xu X., Ding X, & Zhao S. (2023). Downregulation of KLF10 contributes to the regeneration of survived renal tubular cells in cisplatin-induced acute kidney injury via ZBTB7A-KLF10-PTEN axis. Cell Death Discovery, 9, 82.

+ Open protocol

+ Expand

Westerns Blot Analysis of PTEN, p53

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

Total protein was extracted using RIPA buffer (Sigma). The protein concentration was verified by the BCA protein assay kit (Pierce). The proteins were segregated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (Bio-Rad, Hercules, CA) and transferred onto polyvinylidene difluoride membrane. The primary antibodies used were anti-PTEN (1:1,000, ab170941, Abcam, Cambridge, MA), anti-p53 (1:1,000, ab131442, Abcam), and anti-GAPDH (1:10,000, ab181602, Abcam). The membranes were washed and then incubated in goat anti-rabbit horseradish peroxidase‒conjugated secondary antibody (1:2,000) for 2 hours at 37°C. GAPDH was deemed an internal control. An enhanced chemiluminescence kit (Life Technology) used to visualize the immunoblot bands, of which the optical density was analyzed by ImageJ2X software.

+ Open protocol

+ Expand

Western Blot Analysis of Cell Signaling Proteins

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

Antibodies to USP13 (ab99421), NF-kB p65 (ab16502), NF-kB phosph-p65 (ab86299), pan-AKT (ab8805), phosph-AKT (Ser473) (ab8932), phosph-AKT (Thr308) (ab8933), PTEN (ab170941) and GAPDH (ab181602) (all from abcam) were used according to the manufacturers’ protocols. The western blotting analysis was performed as described before. Briefly, equal amounts of protein extracts were separated by 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene fluoride (PVDF) membranes (Millipore, Billerica, MA, USA). The membranes were blocked with Tris-buffered saline plus Tween-20 (TBS-T; 0.1% Tween-20) with 5% (w/v) non-fat dry milk and were then incubated with primary antibodies in TBS-T at 4 °C overnight. After three washes with TBS-T for 15 min each, the membranes were incubated with the appropriate HRP-labeled secondary antibodies for 1 h at 37 °C. The immunobands were visualized using the ECL reagents (Transgen Biotechnology, Beijing, China) on a MicroChemi Chemiluminescent Imaging System (DNR Bio-Imaging Systems, Mahale HaHamisha, Jerusalem, Israel). The densitometric values for each band were calculated by Image J 1.46r software (Wayne Rasband, National institutes of Health, Bethesda, MA, USA), and the statistical analysis were conducted based on the ratios of target protein/GAPDH.

Man X., Piao C., Lin X., Kong C., Cui X, & Jiang Y. (2019). USP13 functions as a tumor suppressor by blocking the NF-kB-mediated PTEN downregulation in human bladder cancer. Journal of Experimental & Clinical Cancer Research : CR, 38, 259.

+ Open protocol

+ Expand

Immunohistochemical Analysis of PTEN in AP Rats

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

All the rats in each group were sacrificed and the pancreas tissues of the AP rats and the control ones were collected. For the tissue fixation, the obtained pancreas tissues were immersed in 4% paraformaldehyde for 12 h followed by dehydration using 30% sucrose solution. Then slices of the pancreas tissues (14 μm) were prepared and co-incubated with primary antibodies against PTEN (dilution, 1 : 50; Abcam, ab170941; rabbit anti-rat) overnight. Subsequently, horseradish peroxidase-conjugated goat anti-rabbit IgGs (dilution, 1 : 1000; Abcam, ab150077) was introduced and incubated with the slices for 1 h. Finally, the slices were treated with diaminobenzidine (DAB, Sigma-Aldrich, St. Louis, MO, USA) before determination under a microscope (OLYMPUS, Tokyo, Japan).

Yan H., Jiang L., Zou H., Chen T., Liang H, & Tang L. (2019). PTEN suppresses the inflammation, viability, and motility of AP-AR42J cells by activating the Wnt/β-catenin pathway. RSC Advances, 9(10), 5460-5469.

+ Open protocol

+ Expand

Protein Expression Analysis by Western Blot

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

Tissues and cells were lysed with RIPA lysis buffer (Beyotime, Shanghai, China). Samples were added to an equal volume of 2 × SDS loading buffer, and centrifuged at 12000 g for 1 min, and 10 μL of sample was added to the sample cell for electrophoretic separation. The membrane was washed with TBS for 5 min after the blots transferred to PVDF membranes, blocked in 5% milk, and then incubated with primary antibodies against PTEN (1:500 dilution, ab170941, Abcam, Cambridge, UK), protein kinase B (AKT; 1:1000 dilution, ab8805, Abcam), phosphorylated AKT (p-AKT; 1:1000 dilution, ab38449, Abcam), acetyl-histone 3 (Ac-H3; 1:500 dilution, 8172T, Cell Signaling Technology, USA), histone 3 (H3; 1:500 dilution, 4499S, Cell Signaling Technology), acetyl-histone 4 (Ac-H4; 1:500 dilution, 13944S, Cell Signaling Technology), histone 4 (H4; 1:500 dilution, 13919S, Cell Signaling Technology). Then, the membranes were incubated with secondary antibody (1:500 dilution, a32733, Abcam). The bands were visualized with ZF-388 gel imaging system (Sanli, China).

Zhang J., Wang P, & Cui Y. (2021). Long noncoding RNA NEAT1 inhibits the acetylation of PTEN through the miR-524-5p /HDAC1 axis to promote the proliferation and invasion of laryngeal cancer cells. Aging (Albany NY), 13(22), 24850-24865.

+ Open protocol

+ Expand

Fluoxetine and Ginsenoside Modulation of LPS-Induced Inflammation

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

Fluoxetine hydrochloride capsule (Flu) was purchased from Patheon France and was packaged by Lilly Suzhou Pharmaceutical Co., Ltd. Ginsenosides Rg1 (GRg1, B21057), and saikosaponin d (Ssd, B20150) were purchased from Yuanye Biotech (Shanghai, China). Lipopolysaccharide (LPS: L3129) was purchased from Sigma–Aldrich. A small-molecule-specific inhibitor of TLR4 signaling (TAK242, A3850) and PI3K/AKT inhibitor (LY294002, A8250) were from Apex Bio (Houston, Texas). TNF-α, IL-6, and IL-1β ELISA kits were purchased from BioProducts MD (Middletown, MD, USA). The antibodies to PI3K (ab178846), p-AKT (ab192623), AKT (ab179463), p-PTEN (ab76431), PTEN (ab170941), p-FOXO1 (ab131339), and FOXO1 (ab52857) were from Abcam, TLR4 antibody (AF7017) was from Affinity Biosciences, the p-PI3K-antibody (#4228) was from Cell signaling, the anti-β-actin (sc-47778) and goat anti-rabbit IgG (sc-516102) were from Santa Cruz Biotechnology.

Xu M., Zhai W., Zhang Y., Pan J., Li J, & Huang S. (2023). Kaixin Jieyu Granule attenuates neuroinflammation-induced depressive-like behavior through TLR4/PI3K/AKT/FOXO1 pathway: a study of network pharmacology and experimental validation. BMC Complementary Medicine and Therapies, 23, 156.

+ Open protocol

+ Expand

Protein Extraction and Western Blotting

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

After the medium was discarded, the cells were lysed with RIPA lysis buffer (Beyotime, Shanghai, China), and the supernatant was gathered after high-speed centrifugation. After quantifying the protein using BCA reagent (Pierce, Rockford, IL, USA), the supernatant was heated in a thermostat water bath at 100°C for 10 min to denature the protein. Thereafter, the total protein (30 µg/lane) underwent SDS-PAGE. Then the protein was transferred to a PVDF membrane (Millipore, Bedford, MA, USA), and blocked in 5% skim milk for 1 h at room temperature. Following that, the membrane was incubated with anti-PTEN antibody (Abcam, ab170941, 1:1000) and anti-GAPDH antibody (Abcam, ab9485, 1:3000) overnight at 4°C. Subsequently, the PVDF membrane was rinsed with TBST solution and incubated with Goat Anti-Rabbit IgG H & L (Abcam, ab125900, 1:1000) for 1 h at room temperature. After the membrane was rinsed with TBST solution again, the protein bands were developed using hypersensitive ECL (Amersham Pharmacia Biotech, Little Chalfont, UK).

Zhang C.C., Li Y., Feng X.Z, & Li D.B. (2021). Circular RNA circ_0001287 inhibits the proliferation, metastasis, and radiosensitivity of non-small cell lung cancer cells by sponging microRNA miR-21 and up-regulating phosphatase and tensin homolog expression. Bioengineered, 12(1), 414-425.

+ 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!