Ab32538

Manufactured by Abcam

Sourced in United States

Ab32538 is a lab equipment product from Abcam. It is designed for use in scientific research applications.

Automatically generated - may contain errors

Lab products found in correlation

Ab17942, by Abcam (2 mentions) Ab32505, by Abcam (2 mentions) Ab81283, by Abcam (2 mentions) Ab8227, by Abcam (2 mentions) Ab184699, by Abcam (2 mentions) Protease and phosphatase inhibitors cocktail, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Euroclone (1 mentions) Supersignal west pico plus chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Ta150041, by OriGene (1 mentions) Anti fgfr1, by Abcam (1 mentions) Non fat dried milk, by Euroclone (1 mentions) Anti pstat3 d3a7, by Cell Signaling Technology (1 mentions) Anti stat3, by Merck Group (1 mentions) β actin, by Merck Group (1 mentions) Bradford assay, by Bio-Rad (1 mentions) Chemiluminescence detection system, by Santa Cruz Biotechnology (1 mentions) Fab3744f, by R&D Systems (1 mentions) Ic0041p, by R&D Systems (1 mentions) Cytofix cytoperm fixation permeabilization solution kit, by BD (1 mentions) Anti β actin antibody, by Merck Group (1 mentions)

7 protocols using ab32538

Western blot analysis of signaling proteins

Cited 1 time

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

Cell pellets were resuspended and lysed in RIPA buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1% Triton X-100, 10% glycerol), complemented with protease and phosphatase inhibitors cocktail (ThermoScientific). Total proteins extracts concentrations were determined through Bradford assay (Bio-Rad). Cytosol and nucleus protein fractions were obtained as previously described [47 (link)].
After 1 h blocking with 5% non-fat dried milk (EuroClone) or bovine serum albumin (SERVA) in Tris-buffered saline with 0.1% Tween (TBS-T), membranes were incubated with primary antibodies at 4 °C overnight. Primary antibodies used: anti-pFGFR1 (06-1433, Millipore), anti-FGFR1 (Abcam ab137765), anti-pSTAT3 (D3A7, Cell Signaling), anti-STAT3 (06596, Millipore), anti-pAKT1 (ab81283; Abcam), anti-AKT1 (ab32505; Abcam), anti-pERK1/2 (ab32538; Abcam), anti-ERK1/2 (ab17942; Abcam), anti t-GFP (TA150041, Origene) and β-actin (Sigma, A5441). After membrane incubation with horseradish-peroxidase-conjugated anti-rabbit secondary antibody (Immuno Reagents), the positive bands were visualized using the ECL kit SuperSignalTM West Pico PLUS Chemiluminescent Substrate (Thermo Scientific) as previously shown [48 (link)].

Cimmino F., Montella A., Tirelli M., Avitabile M., Lasorsa V.A., Visconte F., Cantalupo S., Maiorino T., De Angelis B., Morini M., Castellano A., Locatelli F., Capasso M, & Iolascon A. (2022). FGFR1 is a potential therapeutic target in neuroblastoma. Cancer Cell International, 22, 174.

+ Open protocol

+ Expand

Protein Expression Analysis of GO NPs

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

After treatment with GO NPs treatments, the cells were collected and washed with PBS, and the total proteins were extracted. The membranes were blocked in 5% (w/v) bovine serum albumin (BSA) in TBST was incubated overnight with primary antibodies (β-actin, ERK1/2, and p-ERK1/2) overnight at 4°C. After washing with TBST several times, the membranes were incubated with second antibodies for 60 mins at room temperature. A chemiluminescence detection system (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used to observe bands. The primary antibodies used in this study included β-actin(ab8227, Abcam), ERK1/2(ab184699, Abcam), and pERK1/2(ab32538, Abcam).

Lin L., Zhuang X., Huang R., Song S., Wang Z., Wang S., Cheng L, & Zhu R. (2020). Size-Dependent Effects of Suspended Graphene Oxide Nanoparticles on the Cellular Fate of Mouse Neural Stem Cells. International Journal of Nanomedicine, 15, 1421-1435.

+ Open protocol

+ Expand

Profiling Cellular Signaling Pathways

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

To analyze the activation of ERK1/2 and STAT3 and the expression levels of FPR1 and FPR2, cells were fixed and permeabilized by BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (BD Pharmingen, San Jose, CA, USA). Subsequently, cells were stained with anti-ERK1 (1:100; (phospho T202/Y204) + ERK2 (phospho T185/Y187); ab32538, Abcam)); anti-STAT3 (1:100; phospho Y705 AF4607; R&D Systems, Minneapolis, MN, USA); anti-FPR1 FITC-conjugated (1:50; FAB3744F; R&D Systems); and anti-FPR2 PE-conjugated (1:50; FAB3479P; R&D Systems). Anti-rabbit IgG-FITC (1:200, 656111, Thermo Fisher Scientific) was used as a secondary antibody for ERK1/2 and STAT3 staining. The isotype controls for FPR1 ((anti-mouse IgG2A-FITC control) 1:50; IC003F; R&D Systems) and FPR2 ((anti-mouse IgG2B-PE control) 1:50; IC0041P, R&D Systems) were also used.

Vecchi L., Mota S.T., Zóia M.A., Martins I.C., de Souza J.B., Santos T.G., Beserra A.D., de Andrade V.P., Goulart L.R, & Araújo T.G. (2022). Interleukin-6 Signaling in Triple Negative Breast Cancer Cells Elicits the Annexin A1/Formyl Peptide Receptor 1 Axis and Affects the Tumor Microenvironment. Cells, 11(10), 1705.

+ Open protocol

+ Expand

Immunoblotting for EPHB4, pERK, ERK

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

Total lysates of 50 µg were loaded and run on 12% polyacrylamide gels, which were then blotted onto polyvinylidene difluoride membranes (BioRad). These membranes were incubated with the following antibodies: polyclonal rabbit anti‐EPHB4 (1:500; 20883‐1‐AP, Proteintech); rabbit polyclonal anti‐pErk1/pErk2 (Cat. No ab32538; 1:250 dilution; AbCam) and rabbit polyclonal anti‐Erk1/2 (Cat. No ab17942; 1:1000 dilution; AbCam). An anti‐beta‐actin antibody (1:5000; Sigma) was used in control for the equal loading of the total lysates.

Andolfo I., Lasorsa V.A., Manna F., Rosato B.E., Formicola D., Iolascon A, & Capasso M. (2020). Kinome multigenic panel identified novel druggable EPHB4‐V871I somatic variant in high‐risk neuroblastoma. Journal of Cellular and Molecular Medicine, 24(11), 6459-6471.

+ Open protocol

+ Expand

Selinexor-Induced Tumor Regression Analysis

Cited 1 time

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

KT-10 tumors that regress rapidly after treatment with selinexor, were harvested 24 hours after a single dose of drug (10 mg/kg). Other, less responsive tumors, were harvested 2 hours after dose 6 (MWF dosing) at 10 mg/kg/dose. Tumors were processed for immunoblotting as previously described [30 (link)]. Immunoblots were probed for p53, p21, PARP and cleaved PARP and XPO1/CRM1. Three control and three treated tumors were analyzed for each xenograft line. GAPDH was used as a loading control. KT-10, KT-11, SK-NEP-1, CHLA258, Rh28 and Rh30 tumors were fixed in 10% buffered formalin and paraffin sections were analyzed by H&E as well with the following antibodies: FOXO1 (#2880, Cell Signaling), IKB (ab32518, Abcam), NFKB (sc-8008, Santa Cruz), pRb-phos (ab76298, Abcam), Mcl-1 (sc-819, Santa Cruz), β-catenin (#610154 BD), ERK-phos (ab32538, Abcam), survivin (ab24479, Abcam), and p53 (sc-126, Santa Cruz).

Attiyeh E.F., Maris J.M., Lock R., Reynolds C.P., Kang M.H., Carol H., Gorlick R., Kolb E.A., Keir S.T., Wu J., Landesman Y., Shacham S., Lyalin D., Kurmasheva R.T., Houghton P.J, & Smith M.A. (2015). Pharmacodynamic and genomic markers associated with response to the XPO1/CRM1 inhibitor selinexor (KPT-330): a report from the Pediatric Preclinical Testing Program. Pediatric blood & cancer, 63(2), 276-286.

+ Open protocol

+ Expand

Cell Culture and Signaling Pathway Modulation

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

Cell culture and chemicals. HO-8910, HO-8910pm (a highly metastatic human ovarian cancer) and SKOv3 cell lines were cultured in RPMI-1640 (HyClone, Logan City, UT, USA) medium supplemented with 10% of fetal bovine serum (FBS; gibco, grand Island, NY, USA) and maintained in a humidified incubator at 37˚C and 5% CO 2 . The following antibodies were purchased for western blotting and immunohistochemical staining: anti-AKT (ab32505; Abcam, Cambridge, UK), p70S6K (9202; Cell Signaling Technology, Inc., Danvers, MA, USA), anti-ERK1/2 (AM2189b; Abgent, San Diego, CA, USA), anti-Sp1 (sc14027; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), anti-phospho-AKT (ab81283; Abcam), anti-phsopho-p70S6K (9234; Cell Signaling Technology), anti-phospho-ERK1/2 (ab32538), anti-phospho-Sp1 (T453) and (ab59257) (both from Abcam), anti-phospho-Sp1 (T739) (SAB4504535; Sigma, St. Louis, CA, USA), anti-CD147 (24) , and anti-β-actin (AB10024; Sangon Biotech, Shanghai, China). LY294002 (L9908; Sigma), rapamycin (R706203; Sangon Biotech) and PD98059 (P215; Sigma) were diluted in dimethyl sulfoxide (DMSO).

Zhao J., Ye W., Wu J., Liu L., Yang L., Gao L., Chen B., Zhang F., Yang H, & Li Y. (2015). Sp1-CD147 positive feedback loop promotes the invasion ability of ovarian cancer. Oncology reports, 34(1).

+ Open protocol

+ Expand

Hippocampus and Striatum Protein Analysis

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

The total proteins in the hippocampus and striatum cells were extracted. Primary antibodies CB1 (ab23703, abcam), MEK1 (ab32091, abcam), p-MEK1 (ab214445, abcam), ERK1/2 (ab184699, abcam), p-ERK1/2 (ab32538, abcam) and β-actin (ab8227, abcam) were used. The Western blot procedure is described above.

He X., Yang L., Wang M., Zhuang X., Huang R., Zhu R, & Wang S. (2017). Targeting the Endocannabinoid/CB1 Receptor System For Treating Major Depression Through Antidepressant Activities of Curcumin and Dexanabinol-Loaded Solid Lipid Nanoparticles. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 42(6).

+ 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!