Coronal sections 40 μm-thick containing the sensorimotor cortex were treated with 0.3% H2O2 in PBS to block the endogenous peroxidase activity, followed by treating with blocking solution to reduce the non-specific bindings. The sections were then incubated with primary antibodies against NeuN (1:500; Merck Millipore, Darmstadt, Germany), Foxp1 (1:1,000; Abcam, Cambridge, UK), Cux1 (1:1,000; Santa Cruz Biotechnology, Dallas, TX, USA), GAD65/67 (1:2,000; Sigma, St. Louis, MO, USA), Parvalbumin (Pvalb or PV, 1:3,000; Sigma) and Calretinin (Calb2 or CR, 1:3,000; Merck Millipore) at room temperature overnight. Next, sections were incubated with biotinylated secondary antibodies against mouse or rabbit IgG (1:500; The Jackson ImmunoResearch Laboratories, West Grove, PA, USA) for 2 h at room temperature followed by Vectastain [avidin-biotin complex (ABC) kit, Vector Laboratories, Burlingame, CA, USA] incubation for 1 h. Last, sections were processed with 2 mg/ml of 3,3′-Diaminobenzidine (DAB) with 0.01% H2O2 in PBS and mounted with a glycerol-based aqueous mounting medium.
Foxp1
Manufactured by Abcam
Sourced in United States
FOXP1 is a transcription factor that belongs to the Forkhead box (FOX) family of proteins. It plays a role in the regulation of gene expression and is involved in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Synaptophysin, by Synaptic Systems (2 mentions)
Ankyring, by Merck Group (2 mentions)
Islet 1, by Abcam (2 mentions)
Stmn2, by Novus Biologicals (2 mentions)
Smi 32, by BioLegend (2 mentions)
Synapsin, by Merck Group (2 mentions)
Psd95, by Abcam (2 mentions)
Vectastain avidin biotin complex kit, by Vector Laboratories (1 mentions)
Gad65 67, by Merck Group (1 mentions)
Neuronal nuclei (neun), by Merck Group (1 mentions)
Cleaved caspase 3, by Santa Cruz Biotechnology (1 mentions)
Foxo1 c29h4, by Cell Signaling Technology (1 mentions)
Mum1 mum1p, by Agilent Technologies (1 mentions)
Cd10 56c6, by Leica (1 mentions)
Bcl 6 ln22, by Leica (1 mentions)
Ventana benchmark xt, by Roche (1 mentions)
Cxcr4, by Abcam (1 mentions)
Bcl2 clone 124, by Agilent Technologies (1 mentions)
P53 do 7, by Agilent Technologies (1 mentions)
Cd30 clone berh2, by Agilent Technologies (1 mentions)
10 protocols using foxp1
1
Immunohistochemical Labeling of Cortical Neurons
2
Immunohistochemical Analysis of Tumor Angiogenesis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Six-micrometer sections of formalin-fixed and paraffin-embedded tissue specimens were stained by an established method described previously [18 (link)]. Sections were incubated with antibodies specific for Factor VIII (DAKO, Tokyo, Japan), vascular endothelial growth factor (VEGF) (R & D Systems, Minneapolis, MN), cleaved caspase-3 (Santa Cruz Biotechnology, Santa Cruz, CA), human P450 aromatase (ARK Resource, Kumamoto, Japan), ERα (Santa Cruz Biotechnology) and FOXP1 (Abcam, Tokyo, Japan) at 4°C overnight. Slides were incubated with biotinylated species-specific appropriate secondary antibodies for 30 minutes and exposed to avidin-biotin-peroxidase complex (VECTA Laboratories, Burlingame, CA). Sections were treated with 0.02% DAB as a chromogen and counterstained with hematoxylin. Microvessel density (MVD) was determined as follows. The highly vascularized areas of the tumor stained with an anti-Factor VIII antibody were identified and Factor VIII-positive microvessels were counted within a high-power field (number per 0.75 mm2). Single endothelial cells or clusters of endothelial cells, with or without lumen, were considered individual vessels. MVD was expressed as the vessel number/high-power field in sections. Three fields were counted per animal and the average was taken as the MVD of each tumor.
3
DLBCL Subtyping and FOXO1 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FFPE tissues were subjected to IHC using primary antibodies as follows: CD10 (56C6, Novocastra, Newcastle Upon Tyne, UK), BCL-6 (LN22, Novocastra), MUM1 (MUM1P, Dako, Glostrup, Denmark), GCET1 (polyclonal, Abcam, Cambridge, UK), FOXP1 (polyclonal, Abcam), FOXO1 (C29H4, Cell Signaling, Danvers, MA, USA) and Bim (Y36, Abcam) using the Ventana Benchmark XT automated staining system (Ventana Medical Systems, Tucson, AZ, USA). DLBCL cases were immunohistochemically sub-grouped into GCB or ABC phenotypes according to the Choi algorithms, as previously described [49 (link)]. FOXO1 expression was evaluated according to the staining intensity (0-3) and location (nuclear vs. cytoplasmic).
4
Prognostic Biomarkers in Diffuse Large B-Cell Lymphoma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue microarrays prepared from the diagnostic formalin-fixed, paraffin-embedded (FFPE) tissue blocks of all patients studied were stained with an anti-p63 antibody (4A4, Santa Cruz Biotechnology, Santa Cruz, CA) which can detect all p63 isoforms. Expression levels of p63 were determined by estimating the percentage of p63-positive tumor cells in the tissue array cores. X-tile software and receiver operating characteristic curve analysis by GraphPad Prism 6 Software were used to determine the percentage of p63-positive cells with maximal discriminatory power for the separation of DLBCL patients into 2 different prognostic groups. Evaluation of other biomarkers by immunohistochemistry was also performed on tissue microarrays using corresponding antibodies: p53 (DO-7, Dako, Carpinteria, CA), MDM2 (IF2, Calbiochem, Billerica, MA), p21 (Dako), Bcl-2 (Clone-124, Dako, Carpinteria, CA), Ki-67 (Dako), CD30 (clone BerH2, Dako), Bcl-6 (Dako), FOXP1 (Abcam), IRF4/MUM1 (Dako), CD10 (56C6, Vantana), c-Rel (Dako), and CXCR4 (Abcam, San Francisco, CA). Details of immunohistochemistry procedures and scoring processes have been described previously [38 (link), 44 (link), 55 (link)–58 (link)].
5
Western Blot Analysis of Nuclear Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total cell lysates (60 μg) and nuclear lysates (5 μg), prepared as described42 (link) or by nuclear extraction kit (Panomics, USA), respectively, were subjected to SDS-PAGE and immunoblotting using primary antibodies against PRDM151 (link), actin (1: 5,000 dilution; Sigma), BACH2 (1: 1,000 dilution; abcam), BCL6 (1: 1,000 dilution; Cell Signaling and 1: 1,000; BioLengend), FOXP1 (1: 1,000 dilution; abcam), PAX5 (1: 500 dilution; Santa Cruz), XBP-1 (1: 500 dilution; Santa Cruz), p65 (1: 500 dilution, Santa Cruz), lamin B (1: 400 dilution; Santa Cruz) and GFP (1: 1,000 dilution; Santa Cruz). The detailed procedures, secondary antibodies used and detection of immunoreactive proteins were as described42 (link).
6
Immunostaining of neural markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were washed once with PBS, fixed with 4% PFA for 20 min, washed again in PBS and blocked for 1 h in 0.1% Triton in PBS with 10% donkey serum. Fixed cells were then washed and incubated overnight with primary antibodies at 4◦C. Primary antibody solution was washed and cells were subsequently incubated with secondary antibodies (1:2000, Alexa Fluor, Life Technologies) at room temperature for 1 h, washed with PBS and stained with DAPI. Primary antibodies used: Tuj1 (R&D, MAB1195), Islet1 (Abcam, ab178400), MAP2 (Abcam, ab5392), Synapsin (Millipore, AB1543), SMI-32 (BioLegend, 801,702), Chat (Millipore, AB144P), Foxp1 (Abcam, ab16645), AnkyrinG (Millipore, MABN466), Synaptophysin (Synaptic Systems, 101,004), PSD-95 (Abcam, ab2723), STMN2 (Novus NBP49461). Images were analyzed using FIJI.
7
Immunostaining of neural markers
8
Effector CD8+ T Cell Protein Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For Western analysis, whole-cell protein lysates were obtained from effector CD8+ T cells at 4 time points (0, 3, 5, and 8 days post-stimulation) with lysis buffer Proprep (INTRON Biotechnology) by suspending 106 cells in 10 μl buffer and incubating on ice for 30 minutes in the presence of Halt Protease & Phosphatase inhibitors cocktail (Thermo Scientific). Cell lysates were resolved by 10% Bis-Tris SDS PAGE Gel, transferred onto PVDF membrane (Merck), blocked with 5% skim milk in TBST buffer containing 0.1% Tween-20, and probed with the following mAbs: T-bet (4B10, Santa Cruz), Eomes (Y-20, Santa Cruz), Foxp1 (polyclonal, Abcam), and β-actin (C4, Santa Cruz). Quantification of detected protein was performed with an Intelligent Dark Box unit (LAS-3000; Fujifilm) and normalized for loading with the amount of β-actin (1:5000) detected in each lane.
9
Protein Expression Analysis in HUVECs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein was extracted from HUVECs using RIPA buffer (Cell Signaling Technology, Inc.) and quantified using a BCA kit (Beyotime Institute of Biotechnology). Total protein (30 µg/lane) was separated by SDS-PAGE on 10% gel and transferred to PVDF membranes. The membranes were blocked with 5% BSA (Beyotime Institute of Biotechnology) for 1 h at 37˚C and incubated with primary antibodies (Abcam) overnight at 4˚C. The primary antibodies were DUSP12 (1:1,000; cat. no. ab228987; Abcam), Bcl-2 (1:1,000; cat. no. ab32124; Abcam), Bax (1:1,000; cat. no. 182733; Abcam), Cleaved PARP (1:1,000; cat. no. ab32064; Abcam), PARP (1:1,000; cat. no. ab191217; Abcam), VCAM-1 (1:1,000; cat. no. ab134047; Abcam), ICAM-1 (1:1,000; cat. no. ab282575; Abcam), p-eNOS (1:1,000; cat. no. ab215717; Abcam), eNOS (1:1,000; cat. no. ab300071; Abcam), FOXP1 (1:1,000; cat. no. ab134055; Abcam), MAP3K5 (1:1,000; cat. no. ab45178; Abcam), p-MAP3K5 (1:1,000; cat. no. ab278547; Abcam) or GAPDH (1:1,000; cat. no. ab9485; Abcam). The membranes were then treated with HRP-linked secondary antibodies (goat anti-rabbit; 1:5,000; cat. no. ab6721; Abcam) for 2 h at 37˚C and were visualized using an ECL kit (MilliporeSigma) and semi-quantified using ImageJ software (version 1.42; National Institutes of Health).
10
Immunohistochemistry and Immunofluorescence Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Prior to proceeding with the IHC/IF protocol, a standard process of deparaffinizing sections was performed in xylol and alcohol series. Antigen retrieval was performed by boiling sections in citrate buffer (pH 6,0). After three washes in PBS, blocking solution containing 1–3% BSA and 0.5% Triton X-100 (Sigma-Aldrich, St. Louis, MI, USA) in PBS was applied on the sections for 1–2 h. The blocking solution was replaced with primary antibodies diluted in a blocking solution and incubated overnight at 4 °C. The following antibodies were used: NeuN (Abcam, Cambridge, UK), MAP2 (Sigma-Aldrich, St. Louis, MI, USA), MAP2 (Merck Millipore, Burlington, MA, USA), CUX2 (Abnova, Taipei, Taiwan), CUX2 (Abcam, Cambridge, UK), Reelin (Millipore), DCX (Merck Millipore, Burlington, MA, USA), FOXP1 (Abcam, Cambridge, UK), Neuroserpin (Abcam, Cambridge, UK), TLE4 (Santa Cruz Biotechnology, Dallas, TX, USA), Nurr1 (R&D systems, Minneapolis, MN, USA). After incubation, sections were washed in PBS, and appropriate secondary antibodies (Alexa Fluor, Thermo Fisher Scientific, Waltham, MA, USA) were applied for 2 h at RT. Following three washes in PBS, TrueBlack quencher (Biotium, Fremont, CA, USA) was applied on the sections. Finally, the sections were covered using a mounting medium with DAPI (Vectorlabs, Burlingame, CA, USA). The IHC protocol was implemented as previously described [32 (link)].
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!