The tibial bone GPs (n = 9) were used for the Western blot (WB) analysis. The WB analysis was done according to the method explained by Mehmood et al. [44 (link)]. In short, the GP samples’ homogenization was done in PBS and subjected to a 4 °C cold environment for 2 h. Then centrifugation was done at 14,000 rpm for 10 min and SDS-PAGE was performed on 10% polyacrylamide gel to obtain the equivalent quantity of proteins. The incubation of membranes was done for 1.5 h in 5% skimmed milk and subjected to the primary antibody (rabbit polyclonal anti-AKT, 1:1000; Affinity AF6261 and mouse monoclonal anti-PI3K, 1:1000; BIOSS BSM-33219m) at 4 °C for overnight. Saline with 0.1% Tween 20 was used for 5 min to wash the membranes, and then they were subjected to the secondary antibody for 30 min (goat anti-mouse, 1:3000) at normal temperature. Once again, the washing of membranes was done with saline and images were taken using a standard imaging device luminescent Image Analyzer (CA, USA).
Bsm 33219m
Manufactured by Bioss Antibodies
Sourced in China
The BSM-33219m is a laboratory instrument designed for automated cell counting and analysis. It utilizes advanced imaging and computational techniques to provide precise and reliable cell counts and measurements. The core function of the BSM-33219m is to facilitate accurate and efficient cell enumeration and characterization in a research or clinical setting.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using bsm 33219m
1
Western Blot Analysis of AKT and PI3K
2
Immunohistochemical analysis of cancer biomarkers
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Antibodies of EphA2 (220092), CTNNB1 (R22820) and CD31 (347526) were purchased from ZENBIO (http://www.zen-bio.cn/ http://www.bioss.com.cn/
3
Immunohistochemical Analysis of Breast Cancer
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Resected breast cancer tissues and corresponding para-carcinoma tissues from breast cancer patients who underwent surgical resection were fixed in 4% paraformaldehyde overnight. Then, the tissues were embedded with Paraffin, and cut into 4μm slices. After deparaffinization, dehydration, antigen retrieval and blocking endogenous peroxidase, the sections were incubated with primary antibodies against EphA2 (1:500, ZENBIO, 220092), CTNNB1 (1:500, ZENBIO, R22820) and PIK3R1 (1:500, Bioss, Bsm-33219M) at 4°C overnight. The sections were then incubated with HRP‐conjugated secondary antibodies conjugated with HRP followed by exposing to DAB to visualize signals of the antigen and counterstaining with hematoxylin. The sections were imaged under a microscope (BX43, OLYMPUS). Then, the stained slices were quantitatively analyzed by image-Pro Plus Image analysis system.
4
Renal Protein Expression Analysis
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Renal tissue proteins were extracted, quantified, and visualized using western blotting. First, target proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride (PVDF, G6015-0.45, Servicebio) membranes. The PVDF membranes with bound target proteins were then blocked with 5% skim milk for 2 h. Primary antibodies (dilution 1 : 1000) against p-PI3K (BS-5570R, BIOSS, Beijing, China), PI3K (BSM-33219M, BIOSS), p-AKT (AF0908, Affinity, Changzhou, Jiangsu, China), AKT (3063, Cell Signaling Technology, Danvers, MA, USA), TLR4 (GB11519, Servicebio), and NF-κB (ab216409, Abcam, Cambridge, MA, USA) were used, and the membranes were incubated overnight at 4°C on a shaking bed. After washing the membranes, the secondary antibody (1 : 2,000 dilution) was added and incubated at 37°C for 2 h. After applying the ECL color reagent and performing dark chamber exposure imaging, the gray value of the images was analyzed using ImageJ software.
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