Total protein was extracted from cell homogenates and then immunoblotted with the following antibodies at 4 °C overnight: mouse mAb against human FOXP3 (236A/E7, Abcam, USA) at 1:200, rabbit pAb against human FOXP3 (GTX107737, GeneTex, USA) at 1:800, rabbit mAb against human STUB1 (EPR4447, Abcam, USA) at 1:500, rabbit mAb against ubiquitin (EPR8830, Abcam, USA)at 1:1,000, rabbit mAb against K48-linked ubiquitin (EP8589, Abcam, USA) at 1:1,000, rabbit mAb against Akt(pan) (#4685, CST, USA) at 1:1,000, rabbit mAb against pAkt(S473) (#4060, CST, USA) at 1:1,000, and rabbit mAb against pAkt(T308) (no.13038, CST, USA) at 1:1,000. After the final wash, membranes were incubated with horseradish peroxidase-conjugated secondary immunoglobin G (IgG) antibody (Jackson ImmunoResearch Laboratories, Inc., PA, USA) (1:10,000) at room temperature for 1 h. Membranes were developed using the Enhanced Chemiluminescence Detection Kit (Millipore, Germany).
Gtx107737
Manufactured by GeneTex
Sourced in United States
GTX107737 is a high-performance cell counter that can accurately measure the number and viability of cells in a sample. It utilizes advanced optical detection technology to provide reliable and consistent results.
Automatically generated - may contain errors
Lab products found in correlation
Enhanced chemiluminescence detection kit, by Merck Group (1 mentions)
Epr8830, by Abcam (1 mentions)
Ep8589, by Abcam (1 mentions)
Epr4447, by Abcam (1 mentions)
Novolink polymer detection system, by Leica (1 mentions)
Mach 2 double stain 2, by Biocare Medical (1 mentions)
Vulcan fast red, by Biocare Medical (1 mentions)
Discovery c15, by Olympus (1 mentions)
Alexa fluor 594 conjugated goat anti rabbit igg h l, by ZSGB-BIO (1 mentions)
3 protocols using gtx107737
1
Protein Immunoblotting for FOXP3, STUB1, and Akt
2
Tumor-Infiltrating Immune Cell Analysis
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The levels of CD4, CD8, FOXP3, and CD163 were evaluated in the immune cells located in the invasive tumor area [15 (link)]. CD8 and CD163 were demonstrated by staining methods using antibodies to CD8 (Cell Marque, 108R-14) and CD163 (Biocare Medical, ACR353AK) cells. FOXP3 and CD4 cells were evaluated by the double-staining method using antibodies to CD4 (Biocare Medical, ACI3148) and FOXP3 (Genetex, GTX107737) cells. The MACH 2 Double Stain 2 (Biocare Medical) was utilized for the incubation process. FOXP3 and CD4 cells were stained with Vulcan Fast Red (Biocare Medical) [17 (link)].
Immunohistochemical staining of PD-L1 was performed using a mouse monoclonal primary anti-PD-L1 antibody (clone 22C3; Dako; Agilent Technologies, Inc.). Subsequently, the slides were incubated with Novolink Polymer Detection System (Leica Microsystems) as a secondary antibody (Novocastra). The combined positive score (CPS) was used for evaluating immunohistochemical expression of PD-L1 [19 (link)].
Immunohistochemical staining of PD-L1 was performed using a mouse monoclonal primary anti-PD-L1 antibody (clone 22C3; Dako; Agilent Technologies, Inc.). Subsequently, the slides were incubated with Novolink Polymer Detection System (Leica Microsystems) as a secondary antibody (Novocastra). The combined positive score (CPS) was used for evaluating immunohistochemical expression of PD-L1 [19 (link)].
3
Renal Tissue Immunofluorescence Analysis
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Immuno uorescence stainings of Fractalkine, p-p38, and Foxp3 proteins were performed on 4 µm-thick para nembedded sections of renal tissues after incubation with a primary antibody targeting Fractalkine (DF12376, 1:80; A nity), Foxp3 (GTX107737, 1:100; GENE TEX), or p-p38 (AF4001, 1:80; A nity) overnight at 4 °C. Next, the samples were incubated with Alexa Fluor 594-conjugated goat anti-rabbit IgG (H + L) (ZF0516, 1:100; ZSGB-BIO) for 1 h at 24 °C. Then, the samples were observed with a uorescence microscope (IX71, Olympus, Tokyo, Japan) equipped with ISC capture software, and the images were collected with a CCD camera (Discovery C15, Olympus, Tokyo, Japan).
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