Total cell extracts were prepared in 1× sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) sample loading buffer. Cell fractions were extracted with nuclear and cytoplasm protein extraction kit (Wanleibio, China). Cell proteins were resolved by SDS-PAGE, transferred to a polyvinylidene difluoride membrane, and probed with GAPDH (1:1000, BM3876, Bosterbio, USA), p-PERK, p-IRE1, ATF4, P16 (1:500, bs-3330R, bs-16698R, bs-1531R, bs-20656R, Bioss, China), H3, NRF2 (1:500, D153567, D121053, Sangon Biotech, China), and TNF-α, IL-6, P21, MT1, MT2, HRD1, VCP, P65, p-P65, IKK (1:200, sc-52746, sc-32296, sc-136020, sc-13180, sc-13177, sc-293484, sc-136273, sc-514451, sc-166748, sc-7606, Santa Cruz, USA). Secondary anti-rabbit, anti-mouse antibodies (1:1000, BM2004, BA1001, Bosterbio, USA) conjugated with horseradish peroxidase were used. Detection was performed using a Thermo Scientific Pierce enhanced chemiluminescence western blotting substrate (Thermo Scientific). Results were analyzed by Tanon-410 automatic gel imaging system (Shanghai Tianneng Corporation, China).
Ba1001
Manufactured by Boster Bio
Sourced in United States, China
BA1001 is a pipette designed for precise liquid handling. It features adjustable volume settings and ergonomic design for comfortable use.
Automatically generated - may contain errors
Lab products found in correlation
Sc 13177, by Santa Cruz Biotechnology (1 mentions)
Thermo scientific pierce enhanced chemiluminescence western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Bm3876, by Boster Bio (1 mentions)
Bs 3330r, by Bioss Antibodies (1 mentions)
D121053, by Sangon (1 mentions)
Sc 32296, by Santa Cruz Biotechnology (1 mentions)
Sc 136020, by Santa Cruz Biotechnology (1 mentions)
Sc 514451, by Santa Cruz Biotechnology (1 mentions)
Sc 166748, by Santa Cruz Biotechnology (1 mentions)
Nuclear and cytoplasm protein extraction kit, by Wanlei (1 mentions)
Sc 52746, by Santa Cruz Biotechnology (1 mentions)
Ba1046, by Boster Bio (1 mentions)
Ab14826, by Abcam (1 mentions)
Ar1006, by Boster Bio (1 mentions)
Fv500, by Olympus (1 mentions)
Rabbit anti cd146 antibody, by Abcam (1 mentions)
Ba1003, by Boster Bio (1 mentions)
4 protocols using ba1001
1
Protein Expression Analysis Protocol
2
Immunohistochemical Analysis of LAIR-1 Expression in Tumors
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IHC staining was conducted to examine the altered expression of LAIR-1 in tumors and normal tissues. Formalin-fixed and paraffin-embedded sections were deparaffinized and rehydrated. Antigen retrieval was performed in a microwave at 98°C for 30 min. The sections were treated with 3% hydrogen peroxide for 5 min, and then blocked with 1% bovine serum albumin (AR1006; Boster Biological Technology, Ltd., Wuhan, China) for 30 min. The sides were incubated with monoclonal mouse anti-human LAIR-1 (ab14826; Abcam, Cambridge, UK) at 1:500 dilution at 4°C overnight. Mouse IgG (BA1046; Boster Biological Technology, Ltd.) served as the negative control. The biotinylated anti-mouse secondary antibody (BA1001; Boster Biological Technology, Ltd.) was incubated the following day with the sides for 20 min at room temperature, followed by further treatment with a streptavidin-alkaline phosphatase complex (Boster Biological Technology, Ltd.) for 20 min. The signal was detected using the BCIP/NBT substrate solution (Boster Biological Technology, Ltd.).
3
Immunohistochemical Analysis of Angiogenesis Markers
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Proximal tibias of 1-week-old SD rats were isolated and fixed with 4% formaldehyde. The samples were decalcificated with 12.5% EDTA solution, dehydrated, embedded into paraffin wax and sectioned at 5 mm thickness. Primary mouse anti-CD105 antibody (dilution 1:100) and rabbit anti-CD146 antibody (dilution 1:250) (both from Abcam) were used for immune labeling according to manufacturer's instructions. Then sections were incubated with biotinylated goat anti-mouse or goat anti-rabbit secondary antibodies (#BA1001 or #BA1003, dilution 1:2,000; Boster Biological Technology, Wuhan, China) at RT for 30 min. Reactivity was detected with a diaminobenzidine tetrahydrochloride (DAB) substrate kit (Beyotime Institute of Biotechnology), with hematoxylin as the counterstain. Images were captured by microscopy (FV500; Olympus Corporation, Tokyo, Japan).
4
Immunohistochemical Analysis of Femoral Angiogenesis
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After CT scanning, femurs were decalcified with 10% EDTA solution for 3 weeks and embedded in paraffin wax. The femoral heads were sectioned at 5-μm thickness in the coronal plane. Some of these sections were stained with hematoxylin and eosin (H&E) to evaluate the trabecular structure, while the others were deparaffinized; antigen retrieved; incubated with anti-CD31, anti-VEGF, and anti-VEGFR2 primary antibodies; and then incubated with the corresponding biotinylated goat anti-rabbit (Boster, China, BA1003) and goat anti-mouse (Boster, China, BA1001) secondary antibodies. Sections were colored with DAB and counterstained with hematoxylin. The images of immunohistochemical staining were analyzed using the software Image-Pro Plus. At least five random fields in the femoral head were selected, and the positive staining was quantified based on the integrated option density (IOD) of target proteins. The corresponding area was also measured, and the mean density was defined as the ratio of integrated optical density to the corresponding area.
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