Xuebijing injection (catalog number: z20040033, batch number: 1905061) was manufactured by Tianjin Chase Sun Pharmaceutical Co., Ltd (Tianjin, China). Paeoniflorin (CAS #:23180-57-6), Hydroxysafflor yellow A (CAS #: 78281-02-4), Ferulic acid (CAS # 537- 98-4), and protocatechuic aldehyde (CAS #: 139-85-5) were purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). Mouse NT-proBNP ELISA (Cat#: ZC-37812-96T) kit, DCFH-DA (Cat#:CA1410-100T), RIPA tissue/cell lysate (Cat#: R0020), SDS-PAGE gel kit, and Rainbow 245 Spectrum Marker (Cat#:PR 1920), Mouse IL-6 ELISA kit (Cat#:ZC-37988-96T), Mouse IL-1β ELISA kit (Cat#:ZC-37974-96T), and EasySee Western Blot Kit (Lot#:O30807) were purchased from Solarbio (Beijing, China). The primary antibody of rabbit anti-CXCL2 (bs-1162R) was purchased from Bioss Inc., (Beijing, China) and GAPDH, 1:4,000 (14C10) was from Cell Signaling Technology (Beverly, MA, United States). The secondary antibody Goat anti-Rabbit IgG (1:4,000) (ZB-2301) was purchased from ZS bio (Beijing, China). 2, 2, 2-Tribromoalcohol (Cat#:T48402) and Bacterial lipopolysaccharide (Cat#: L2880-10 MG) were purchased from Sigma Aldrich (St. Louis, MO, United States). Rhod2-AM (Cat#: zy0129) and Anti-GRP78 (BiP) antibody (ab21685) were purchased from Abcam (Cambridge, MA, United States).
Ripa tissue cell lysate
Manufactured by Solarbio
Sourced in China
RIPA tissue/cell lysate is a buffer solution used to extract and solubilize proteins from biological samples, such as cells or tissues. It is designed to disrupt cell membranes and facilitate the release of proteins for further analysis or purification.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay kit, by Beyotime (2 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Paeoniflorin, by Yuanye Bio-Technology (1 mentions)
Protocatechuic aldehyde, by Yuanye Bio-Technology (1 mentions)
Dcfh da, by Solarbio (1 mentions)
Mouse il 6 elisa kit, by Solarbio (1 mentions)
Mouse il 1β elisa kit, by Solarbio (1 mentions)
Bacterial lipopolysaccharide, by Merck Group (1 mentions)
Hydroxysafflor yellow a, by Yuanye Bio-Technology (1 mentions)
Ferulic acid, by Yuanye Bio-Technology (1 mentions)
Rhod 2 am, by Abcam (1 mentions)
Sds page gel kit, by Solarbio (1 mentions)
Amersham hybond, by GE Healthcare (1 mentions)
Sds page protein loading buffer 5x, by Beyotime (1 mentions)
Beyoecl star, by Beyotime (1 mentions)
Hybond, by Cytiva (1 mentions)
Bca protein assay kit, by CWBIO (1 mentions)
Sybr premix ex taq kit, by Takara Bio (1 mentions)
Anti phospho mtor ser2448, by Cell Signaling Technology (1 mentions)
Sybr premix ex taq, by Takara Bio (1 mentions)
5 protocols using ripa tissue cell lysate
1
Xuebijing Injection Protocol for Sepsis
2
Protein Extraction and Western Blot Analysis
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Eight tissue specimen pairs were cut into pieces and placed in 1% PMSF and RIPA tissue/cell lysate (Solarbio, China) on ice for 30 minutes. The lysed tissue was then centrifuged at 12,000 rpm at 4°C for 15 minutes. The supernatant was removed and the protein concentration was determined using the BCA Protein Assay Kit (Beyotime, China). SDS-PAGE protein loading buffer (5X) (Beyotime, China) was added and the mixture was boiled for 10 minutes. The protein (30 μg) was added to a prepared 12% SDS-PAGE gel for electrophoretic separation and transferred to a 0.45 μm PVDF membrane (Amersham Hybond, GE Healthcare). The membrane was blocked with a rapid blocking solution for 30 minutes and incubated with EPHB2 (1:1000, AF5246, Affinity), SLC6A1 (1:1000, DF4510, Affinity), PPP1R17 (1:1000, PA5-61599, Invitrogen), PPARGC1A (1:1000, FNab06351, FineTest), or GAPDH (1:1000, ab181602; Abcam) antibodies overnight on a 4°C shaker and washed three times with TBST (0.1% Tween-20) for 10 minutes. The membrane was then incubated with goat anti-rabbit IgG (H + L) HRP (1:3000, S0001; affinity) for 1 hour, washed, and developed using BeyoECL Star (Beyotime, China).
3
Comprehensive Protein Extraction and Analysis
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Total cell proteins were extracted by RIPA tissue/cell lysate (Solarbio, China). We used a BCA Protein Assay Kit (CWBIO, China) to assess the total protein concentration. To evaluate the expression of protein, we used 10% SDS‒PAGE to isolate the protein samples and transferred the samples to polyvinylidene fluoride (PVDF) membranes. The membranes were incubated with 5% BSA sealer solution for at least 2 hr. Next, the membranes were incubated with primary antibody for 24 hr and then with secondary antibodies for 1–2 hr. Finally, the membranes were developed using Super ECL Plus (UElandy, China). ImageJ software was used for quantitative determination of each strip.
4
Evaluating Dendrobium Polysaccharide's Impact
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Dendrobium officinale polysaccharide (VTY24621, Clara Reagent Grade, 98%) was purchased from Dehang Wuzhou (Beijing, China). Primary antibodies PI3 Kinase p85 (Cat#4257), anti−Phospho-AKT (ser473) (Cat#4060), and anti−Phospho-mTOR (ser2448) (Cat# 5536) were purchased from Cell Signaling Technology (USA). Anti-GPR41 (Cat#AF9057) and anti-GAPDH (Cat#AF7021) were purchased from Affinity Biosciences Technology (Jiangsu, China). SYBR Premix EX Taq™ (RR047A) and SYBR Premix EX Taq™ Kit (RR820A) were purchased from TaKaRa (Dalian, China), and PCR primers were purchased from Gencreate (Wuhan, China). RIPA tissue/cell lysate was purchased from Solarbio Life Sciences (Beijing, China). The ELISA kits [testosterone (E05101m), insulin (E05070r), polypeptide YY (E13432r), and estradiol (E05110r)] were purchased from CUSABIO (Wuhan, China). Metformin hydrochloride (H11021518) was purchased from Beijing Jingfeng Pharmaceutical Group Co., Ltd. (Beijing, China).
5
Quantitative Western Blot Analysis
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The proteins were extracted from brain tissues with high-efficiency RIPA tissue/cell lysate and PMSF (Solarbio, Beijing, China). The protein concentrations were estimated with a BCA protein assay kit (Beyotime Biotechnology, Shanghai, China). In each well, 40 μg of protein were loaded and separated by electrophoresis at 120 V in precast 4–20% polyacrylamide gels (GenScript Biotech, Nanjing, China) and transferred to a nitrocellulose membrane (Millipore, Billerica, MA).
After 1 h at 37 °C in a blocking solution (Beyotime Biotechnology, Shanghai, China), monoclonal or polyclonal antibodies were added to the membrane for overnight incubation at 4 °C. Primary antibodies against p16INK4a, PCNA, Iba1 and PSD95 were from Abcam (Cambridge, UK). Antibodies against β-actin were from Cell Signaling Technology (1:1000; CST, Danvers, MA). On the next day, goat anti-rabbit IgG antibody (1:5000; Abcam, Cambridge, UK, red fluorescence) was added to the membrane at 37 °C for 1 h away from light. The Odyssey imaging system and image analysis software (LI-COR Biosciences, Lincoln, NE) were used to quantify the amount of target protein by grey scanning using β-actin as an endogenous reference protein (Table 2 ).
After 1 h at 37 °C in a blocking solution (Beyotime Biotechnology, Shanghai, China), monoclonal or polyclonal antibodies were added to the membrane for overnight incubation at 4 °C. Primary antibodies against p16INK4a, PCNA, Iba1 and PSD95 were from Abcam (Cambridge, UK). Antibodies against β-actin were from Cell Signaling Technology (1:1000; CST, Danvers, MA). On the next day, goat anti-rabbit IgG antibody (1:5000; Abcam, Cambridge, UK, red fluorescence) was added to the membrane at 37 °C for 1 h away from light. The Odyssey imaging system and image analysis software (LI-COR Biosciences, Lincoln, NE) were used to quantify the amount of target protein by grey scanning using β-actin as an endogenous reference protein (
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