The products of anti-Nrf2 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), anti-HO-1 (StressMarq Biosciences Inc., Cadboro Bay, VIC, Canada), anti-Bcl-xL (BioVision, Inc., Milpitas, CA, USA), anti-phospho Akt (Cell Signaling Technology Japan, Inc., Tokyo, Japan), and anti-β-actin (Wako Pure Chemical Industries, Ltd., Osaka, Japan) were used for detecting protein expression.
Anti bcl xl
Manufactured by Abcam
Sourced in United States, United Kingdom, Canada
Anti-Bcl-xL is a primary antibody that recognizes the Bcl-xL protein, a member of the Bcl-2 family of proteins involved in the regulation of apoptosis (programmed cell death). This antibody can be used in various laboratory techniques to detect and study the Bcl-xL protein.
Automatically generated - may contain errors
Lab products found in correlation
Anti bcl 2, by Abcam (14 mentions)
Anti bax, by Abcam (12 mentions)
Anti mcl 1, by Abcam (6 mentions)
Anti β actin, by Abcam (4 mentions)
Anti cytochrome c, by Abcam (3 mentions)
Anti bad, by Abcam (3 mentions)
Anti survivin, by Abcam (3 mentions)
Anti caspase 3, by Cell Signaling Technology (3 mentions)
Anti puma, by Santa Cruz Biotechnology (3 mentions)
Anti p53, by Abcam (3 mentions)
Anti nrf2, by Santa Cruz Biotechnology (2 mentions)
Anti β actin, by Fujifilm (2 mentions)
Anti ho 1, by StressMarq Biosciences (2 mentions)
Anti phospho akt, by Cell Signaling Technology (2 mentions)
Anti flag, by Merck Group (2 mentions)
Chemidoc mp, by Bio-Rad (2 mentions)
Bovine serum albumin (bsa), by Abcam (2 mentions)
Bradford reagent, by Abcam (2 mentions)
Anti β actin, by Proteintech (2 mentions)
Anti bcl 2, by Cell Signaling Technology (2 mentions)
34 protocols using anti bcl xl
1
Western Blot Analysis of Redox Regulators
2
Fucoxanthin-Induced Antioxidant Response
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Caspase-Glo 3/7, pGL4.37 [luc/ARE/Hygro], and FuGENE 6 plasmid were obtained from the Promega Corporation (Madison, WI, USA). The products of anti-Nrf2 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), anti-HO-1 (StressMarq Biosciences Inc., Victoria, BC, Canada), anti-Bcl-xL (BioVision, Inc., Milpitas, CA, USA), anti-phospho Akt (Cell Signaling Technology Japan, Inc., Tokyo, Japan), and anti-β-actin (Wako Pure Chemical Industries, Ltd., Osaka, Japan) were used for detecting the protein expression. 3-(4,5-Dimethyl-2-thiazlyl)-2,5-diphenyltetrazolium bromide (MTT) and fluorescein were obtained from Nacalai Tesque, Inc. (Kyoto, Japan). 5, 5′-Dimethyl pyrolline N-oxide (DMPO) as a spin trap reagent was purchased from Dojindo Laboratories (Kumamoto, Japan). 2,2′-Aazobis (2-methylpropionamidine) dihydrochloride (AAPH) was purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Fucoxanthin (Fx) from Nemacystus decipiens and its deacetylated, fucoxanthinol (FxOH) were obtained from South Product, Ltd. (Okinawa, Japan).
3
Doxycycline-Induced Apoptosis Analysis
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Cells were harvested 12 h after doxycycline treatment and lysed in RIPA buffer (Thermo Fisher Scientific) supplemented with a protease inhibitor cocktail (Quartett) and a phosphatase inhibitor cocktail (Sigma). For BIM detection, the pan-caspase inhibitor Z-VAD-FMK (MedChemExpress) was added 30 min prior to the addition of doxycycline. The cell lysates were then incubated for 1 h on ice and centrifuged at 15,000g for 30 min at 4 °C. Total protein concentrations for the whole-cell lysates were measured using a BCA protein assay kit (Thermo Fisher Scientific). Equal amounts of the lysates were resolved on SDS-PAGE gels and transferred to PVDF membranes. These were then incubated with primary antibody overnight at 4 °C before being washed three times with TBS-T buffer and incubated with a secondary antibody for 1 h at room temperature. Chemiluminescence was generated with ECL Western Blotting Substrate (Thermo Fisher Scientific) and detected using a ChemiDoc MP (Bio-Rad). All images were analyzed using Image Lab (Bio-Rad). The antibodies used for the western blots were: anti-FLAG (Sigma, F1804), anti-BCL-xL (Abcam, ab32370), anti-MCL-1 (Abcam, ab32087), anti-γ-H2A.X (Abcam, ab81299), anti-PARP (Abcam, 191217), anti-β-actin (Abcam, ab6276), HRP-conjugated anti-rabbit IgG (Abcam, ab205718), and HRP-conjugated anti-mouse IgG (Abcam, ab6728).
4
Apoptotic Effects of AEE Crystal
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AEE transparent crystal with the purity of 99.5% by RE-HPLC was prepared in Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS. H2O2 solution (cat number: 323381), dimethyl sulfoxide (DMSO), reactive oxygen species assay kit, and trypsin-EDTA were supplied by Sigma (St. Louis, MO). Cell Counting Kit-8 (CCK-8) was from MedChemExpress (NJ, USA), DMEM/F12 (1 : 1), and fetal bovine serum was from Gibco (NY, USA). LysoTracker Red probe, Hoechst 33342 staining solution, cellular glutathione peroxidase assay kit, Cu/Zn-SOD and Mn-SOD assay kit, and mitochondrial membrane potential assay kit were purchased from Beyotime (Shanghai, China). MitoTracker Red CMXRos probe was from Thermo Scientific (MA, USA). Anti-Bid cleavage site, Anti-Bax, Anti-Bcl2, Anti-Bcl-XL, anti-Caspase3 (Cas3), anti-cytochrome C, and anti-Cathepsin D (CTSD) were purchased from Abcam (MA, USA) and the CTSD activity assay kit was from BioVision (CA, USA). An Annexin V/FITC apoptosis detection kit was from BD Biosciences (NY, USA). A Caspase-3 activity assay kit was from Cell Signaling Technology (MA, USA).
5
Comprehensive Immune Profiling of CAR-T Cells
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The following mAbs were used in this study; anti‐CD8α, anti‐TIGIT, anti‐CD107a, and anti‐Bcl‐2 (BioLegend), anti‐CD90.2, anti‐PD‐1, anti‐LAG‐3, anti‐TIM‐3, and anti‐Vα8.3 mAb (Thermo Fisher Scientific), antihuman CD20 (BD Biosciences), anti‐Bcl‐xL (Abcam), and anti‐Bim (CST). Annexin V (BD Biosciences) and Zombie Yellow viability dye (BioLegend) were also used. Biotinylated recombinant protein L (GenScript) was used to detect CAR‐T cells, as previously reported.16 Intracellular protein staining buffer set (Thermo Fisher Scientific) was also used in some experiments. APC‐conjugated antihuman IgG mAb (BioLegend) was used to detect the binding of mouse PD‐L1‐human Fc fusion protein (R&D Systems). Antimouse CD16/CD32 mAb was used for blockade of non‐specific binding of mAb to Fcγ receptors. Flow cytometric data were acquired by EC800 (SONY) or BD LSRFortessa X‐20 cell analyzer (BD Biosciences), and analyzed using FlowJo software (FlowJo, LLC).
6
Silymarin-Based Hepatoprotective Assay
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Silymarin, TAA, BSA, Bradford reagent, anti-Bcl-2, anti- Bcl-xL, anti-Bad, and anti-Bax antibodies were purchased from Abcam (UK). Other antibodies were purchased from Sigma-Aldrich Chemical Company (St. Louis, MO, USA). Kits for measurement of blood glucose and LDH were purchased from Span Diagnostic Ltd., India. All other chemicals were bought from Sisco Research Laboratory, India.
7
Western Blotting Analysis of Apoptosis and Inflammation Signaling
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Western blotting analysis was carried out as previously described [25 (link)]. The following antibodies were used: anti-Bcl-2 (#ab182858), anti-Bax (#ab182733), anti-Bcl-xl (#ab32370), anti-p-P65 (#ab76302), anti-P65 (#ab32536), anti-p-IκBα (#ab133462), anti-IκBα (#ab32518) (Abcam Cambridge, MA, USA), anti-FXR (#sc-25309), anti-TLR4 (#sc-293072) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-p-ERK1/2 (#4370), anti-ERK1/2 (#4695), anti-p-P38 (#4511), anti-P38 (#8690), anti-p-JNK1/2 (#4668), anti-JNK1/2 (#9252), anti-CCL2 (#66272-1-Ig), and anti-β-actin (#66009-1-Ig) (Proteintech, Wuhan, China).
8
Protein Expression Analysis Protocol
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Cells were lysed in cell lysis buffer (50 mM Tris pH 8.0, 150 mM NaCl, 10% glycerol, 1 mM EDTA, 50 mM NaF, and 0.1% NP-40) supplemented with protease and phosphatase inhibitors. Total protein extracts of tissues were generated in radioimmunoprecipitation (RIPA) assay lysis buffer containing complete protease and phosphatase inhibitor cocktail. Protein content was quantified using the Pierce BCA Protein Assay Kit (#23225, Thermo Fisher Scientific). The proteins were separated by SDS-PAGE, electrophoretically transferred to polyvinylidene difluoride (PVDF) blotting membranes (#LC2002, Thermo Fisher Scientific), and subjected to routine immunoblot analysis with primary antibodies as follows: anti-Bcl-2 (#15071), anti-Mcl-1 (#94296), anti-VHL (#68547), anti-CRBN (#71810), and anti-Sufu (#2522) were obtained from Cell Signaling Technology (Danvers, Massachusetts, USA); anti-Bcl-xL (#ab32370), anti-Gli1 (#ab134906), and anti-Smoothened (#ab236465) were obtained from Abcam (Cambridge, CB2 0AX, UK); anti-GAPDH (#60004-1-Ig) and anti-β-tubulin (#10094-1-AP) were purchased from Proteintech Group, Inc. (Rosemont, IL 60018, USA). The results were quantified by densitometry using ImageJ software (NIH).
9
Quantitative Immunoblotting for Signaling Proteins
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The following reagents and materials were used: anti-ALDH1B1 (ref. sc-374090) was purchased from Santa Cruz; anti-PI16 (ref. PAS 31882) from Thermo; anti-P70 S6K beta (ref: ab184551) from Abcam; and anti-Bcl-xL (ref. 2764), anti-pAkt (Ser473) (ref. 4060), anti-Akt (ref. 4685), anti-pMEK1/2 (Ser217/221) (ref. 9154), anti-MEK1/2 (ref. 9126), anti-pERK1/2 (Thr202/Tyr204) (ref. 4370), anti-ERK1/2 (ref. 9102), anti-pPKA (Thr197) (ref. 5661), anti-PKA (ref. 4782), anti-pSEK1/MKK4 (Ser257/Thr261) (ref. 9156), anti-SEK1/MKK4 (ref. 9152), anti pSAPK/JNK (Thr183/Tyr185) (ref. 9255), anti pSAPK/JNK (ref. 9252S), anti pMKK3-6 (Thr183/Tyr185) (ref. 9231), anti MKK3 (ref. 5674), anti p-p38 MAPK (Thr180/Tyr182) (ref. 4511), anti p38 MAPK (ref. 9212) were purchased from Cell Signaling. Electrophoresis reagents were purchased from Biorad and trypsin from Promega.
10
Tumor Protein Expression Analysis
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Cells treated with agents were lysed with RIPA buffer, and western blotting was carried out as previously reported.7 (link), 21 (link) In the molecular analysis of tumor samples in vivo, single‐cell suspensions were prepared using the gentleMACS Octo Dissociator (Miltenyi Biotec). After tumors had been transformed to single cells, the cells obtained were treated as described above. Primary Abs were rabbit anti‐survivin (R&D Systems), anti‐Bcl‐2 and PD‐L1 (Abcam), anti‐Bcl‐xL, phosphorylated ERK and Akt, and anti‐total ERK and Akt (Cell Signaling Technology), and mouse anti‐β‐actin (Sigma‐Aldrich) Abs.
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