Mouse polyclonal antibodies to TRIP-Br2 were raised against the recombinant protein of human TRIP-Br2 as previously described8 (link). Anti-BiP (#3177), anti-CHOP (#2895), and anti-cleaved caspase 3 (#9661) antibodies were obtained from Cell Signaling. Anti-ATGL (55190-1-AP) and anti-β-actin (66009-I-Ig) antibodies were from Proteintech. Anti-UCP1 (ab10983) antibody was from Abcam.
Anti β actin 66009 1 ig
Manufactured by Proteintech
Sourced in China, United States
Anti-β-actin (66009-1-Ig) is a primary antibody that recognizes the β-actin protein. This antibody is commonly used in Western blotting and immunofluorescence applications to detect and quantify the expression of β-actin, a housekeeping gene that is ubiquitously expressed in eukaryotic cells.
Automatically generated - may contain errors
Lab products found in correlation
Anti gapdh 60004 1 ig, by Proteintech (3 mentions)
Anti nlrp3 ag 20b 0014, by Adipogen (2 mentions)
Anti akt, by Cell Signaling Technology (2 mentions)
Anti p akt, by Cell Signaling Technology (2 mentions)
Ripa buffer, by Beyotime (2 mentions)
Ecl kit, by Thermo Fisher Scientific (2 mentions)
Anti cleaved caspase 3 9661, by Cell Signaling Technology (1 mentions)
Anti p27 sc 528, by Santa Cruz Biotechnology (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Anti nrf2 16396 1 ap, by Proteintech (1 mentions)
Nigericin, by Merck Group (1 mentions)
Ly6g bv421, by BioLegend (1 mentions)
Anti mouse caspase 1 p20 ag 20b 0042, by Adipogen (1 mentions)
Cd11b percp cy5, by BD (1 mentions)
Cd19 fitc, by Thermo Fisher Scientific (1 mentions)
Cd45 bv510, by BioLegend (1 mentions)
Cd11c percp cy5, by BioLegend (1 mentions)
Ly6g pe, by BioLegend (1 mentions)
Cd45 pe, by Thermo Fisher Scientific (1 mentions)
Cd11b pe cy7, by Thermo Fisher Scientific (1 mentions)
24 protocols using anti β actin 66009 1 ig
1
Western Blot for TRIP-Br2, ER Stress, and Apoptosis
2
Anticancer Drug NSC139021 Characterization
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NSC139021(HY-112158) was purchased from MedChemExpress (Monmouth Junction, NJ, USA) and was solubilized in dimethylsulfoxide (Sigma-Aldrich, St. Louis, MO, USA) at 20 mM and 75 mg/mL stock solution. Antibodies were as follows: anti-Cyclin A (sc-751), anti-CyclinB1 (sc-245), anti-Cyclin E (sc-25303), anti-Skp2 (sc-7164), anti-p-Rb (Ser807) (sc-293117), and anti-p27 (sc-528) were purchased from Santa Cruz Biotechnologies (Dallas, DE, USA); anti-RIOK1 (17222-1-AP), anti-RIOK3 (13593-1-AP), anti-GAPDH (60004-1-Ig), anti-p21 (10355-1-AP), and anti-β-actin (66009-I-Ig) were purchased from Proteintech (Wuhan, China); anti-p53 (2524S), anti-caspase-3 (9662S) were purchased from Cell Signaling Technology (Boston, MA, USA); anti-Bcl-2 (A19693) and anti-Bax (A19684) were purchased from ABclonal (Wuhan, China); anti-RIOK2 (HPA005681) was from Sigma-Aldrich (St. Louis, MO, USA); homemade polyclonal antibody of CDK2 was the gift from Dr. Hui Zhang (Department of Chemistry and Biochemistry, University of Nevada, Las Vegas, NV 89154, USA). Cell Counting Kit-8 (CCK-8) (BA00208) was obtained from Bioss (Beijing, China), and Annexin-V-FITC/PI kit (40302ES60) was purchased from YEASEN (Shanghai, China).
3
Cigarette Smoke-Induced Oxidative Stress
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Cigarette smoke extraction (CSE) was prepared as previously described [15 (link)]. Antibodies against β-actin (anti-β-actin, 66009-1-Ig), TXNRD1 (anti-TXNRD1, 11117-1-AP), and Nrf2 (anti-Nrf2, 16396-1-ap) were purchased from Proteintech. HO-1 (anti-HO-1, GB11845) was purchased from Servicebio. Auranofin (Ridaura, SKF-39162), the inhibitor of thioredoxin reductase (TXNRD1), was purchased from Selleck.cn.
4
Inflammasome Activation Assay Protocol
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HDM was from Greer Laboratories. Anti-mouse IL-1β (p17) (AF-401-NA) was from R&D. Anti-mouse caspase-1 (p20) (AG-20B-0042) and anti-NLRP3 (AG-20B-0014) antibodies were from Adipogen. Anti-ASC (67824) antibody was from Cell Signaling Technology. Anti-β-actin (66009-1-Ig) was from Proteintech.
Anti-mouse antibodies used for flow cytometry were: CD3-FITC (BD, 553062, 145-2C11), CD19-FITC (eBioscience, 11-0193-82, 1D3), Ly-6G-PE (Biolegend, 127608, 1A8), CD45-PE (eBioscience, 12-0451-81, 30-F11), CD11c-PerCP-Cy5.5 (Biolegend, 117328, N418), CD11b-PerCP-Cy5.5 (BD, 550993, M1/70), CD11b-PE-Cy7 (eBioscience, 25-0112-82, M1/70), CD3e-PE-Cy7 (BD, 552774, 145-2C11), CD19-PE-Cy7 (Biolegend, 115520, 6D5), SiglecF-Alexa Fluor 647 (BD, 562680, E50-2440), MHCII-APC-eFluor 780 (eBioscience, 47-5321-82, M5/114.15.2), Ly-6G-BV421 (Biolegend, 127628, 1A8), CD45-BV510 (Biolegend, 103137, 30-F11), CD11c-BV510 (Biolegend, 117338, N418). Ultrapure LPS was obtained from Invitrogen. Nigericin was obtained from Sigma-Aldrich. RRx-001 (S8405) was from Selleck.
Anti-mouse antibodies used for flow cytometry were: CD3-FITC (BD, 553062, 145-2C11), CD19-FITC (eBioscience, 11-0193-82, 1D3), Ly-6G-PE (Biolegend, 127608, 1A8), CD45-PE (eBioscience, 12-0451-81, 30-F11), CD11c-PerCP-Cy5.5 (Biolegend, 117328, N418), CD11b-PerCP-Cy5.5 (BD, 550993, M1/70), CD11b-PE-Cy7 (eBioscience, 25-0112-82, M1/70), CD3e-PE-Cy7 (BD, 552774, 145-2C11), CD19-PE-Cy7 (Biolegend, 115520, 6D5), SiglecF-Alexa Fluor 647 (BD, 562680, E50-2440), MHCII-APC-eFluor 780 (eBioscience, 47-5321-82, M5/114.15.2), Ly-6G-BV421 (Biolegend, 127628, 1A8), CD45-BV510 (Biolegend, 103137, 30-F11), CD11c-BV510 (Biolegend, 117338, N418). Ultrapure LPS was obtained from Invitrogen. Nigericin was obtained from Sigma-Aldrich. RRx-001 (S8405) was from Selleck.
5
Immunoblotting of JAK2 and STAT Signaling
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Immunoblotting was performed as previously described (20 (link)). For examination of the protein involved in pro-survival signaling pathway, cell lysis from empty-, JAK2-, or JAK2V617F-expressing macrophages were extracted and subpackaged equally for three groups (each group included empty, JAK2 and JAK2V617F lysates), followed by separation on 8% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel. After semi-dry transfer, the membranes were sequentially probed with the indicated antibodies. Anti-p-JAK2 (T1007/1008) (#3776), anti-JAK2 (D2E12) (#3230), anti-p-STAT1 (T701) (#7649), anti-STAT1 (D1K9Y) (#14994), anti-p-STAT3 (Tyr705, D3A7) (#9145), anti-STAT3 (D3Z2G) (#12640), anti-p-STAT5 (Tyr694, C11C5) (#9359), anti-STAT5 (D206Y) -(#94205), anti-p-p38 (Thr180/Tyr182, D3F9) (#4511), anti-p38 (D13E1) (#8690), anti-p-AKT (Ser473, D9E) (#10019), anti-AKT (#9272), anti-p-JNK (Thr183/Tyr185, 81E11) (#4668), and anti-JNK (#9252) antibodies were purchased from Cell Signaling Technology. Anti-HK1 (19662-1-AP), anti-HK2 (22029-1-AP), anti-PKM1 (15821-1-AP), anti-flag (Sigma), and anti-β-actin (66009-1-Ig) antibodies were purchased from ProteinTech.
6
Quantitative Analysis of Protein Signaling
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Cells were seeded at 4.5 × 105 cells/dish in 100 mm dishes containing 15 mL of medium and cultured for 48 h. Then, the cells were incubated for 24 h with a fresh medium containing GEM or NANV, or both. Western blot was performed as described previously [30 (link)]. Primary antibodies used are as follows: anti-β-actin (66009-1-Ig) from Proteintech; anti-phospho-Ser240/244-S6 ribosomal protein (5364), anti-S6 ribosomal protein (2217), anti-phospho-Ser51-eIF2α (3398), anti-eIF2α (5324), anti-phospho-Thr37/46-4EBP1 (2855), and anti-4EBP1 (9452) from Cell Signaling Technology.
7
Western Blot Analysis of F11 and PSRC1
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Cell lysates were prepared in RIPA buffer [10% SDS, 20% NP-40, 0.5M EDTA, 5M NaCl, and 1.25M Tris-Cl (pH 8.0)] supplemented with proteasome inhibitors. Equal amounts of protein were separated on SDS-PAGE gels and then transferred to PVDF membranes (Amersham, GE, USA). Membranes were incubated with specific primary antibodies: anti-F11 (45895-1; Signalway Antibody); anti-PSRC1 (32879-1; Signalway Antibody). Anti-β-actin (66009-1-Ig; Proteintech) served as the loading control. HRP-conjugated anti-mouse or anti-rabbit secondary antibodies were used, and signals were detected using Western ECL substrate (Beyotime).
8
Quantitative Protein Expression Analysis
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Western blot was performed as previously described (Jian et al., 2011) using the following antibodies: anti-K17 (ab53707; Abcam), anti-FASN (ab128856; Abcam), anti-SREBP-1 (PA1-337; Invitrogen), anti-PPARγ (81B8; Cell Signaling Technology), anti-β-tubulin (10068-1-AP; Proteintech), anti-β-actin (66009-1-Ig, Proteintech), anti-GAPDH (60004--1-Ig; Proteintech), and anti-lamin-A/C (sc-7292; Santa Cruz Biotechnology). Band intensities were quantified using Image Lab version 5.2.1 (Bio-Rad). Relative band intensities were normalized to that of the loading control.
9
Investigating NLRP3 Inflammasome Activation
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Lipopolysaccharides (LPS, #L2880), adenosine triphosphate (ATP, #A3377), and aluminum potassium sulfate dodecahydrate (Alum, #237086) were purchased from Sigma (St. Louis, MO, USA). COS was bought from Abphyto Biotech Co., Ltd. (Chengdu, China). MitoSOX (#M36008) was obtained from Invitrogen (Carlsbad, CA, USA). MitoTracker (#C1049B) and protein A + G agarose (#P2012) were supplied by Beyotime (Shanghai, China). Anti-NLRP3 (#AG-20B-0014) and anti-mouse caspase-1 (#AG-20B-0042) antibodies were purchased from Adipogen (San Diego, CA, USA). Anti-NEK7 (#ab133514) and anti-GSDMD (#ab209845) antibodies were obtained from Abcam (Cambridge, UK). Anti-mouse IL-1β (#AF-401-NA) antibody was from R&D Systems (Minneapolis, MN, USA). Anti-Flag (#20543-1-AP), anti-HA (#66006-2-Ig), and anti-β-actin (#66009-1-Ig) antibodies were purchased from Proteintech (Rosemont, IL, USA).
10
Immunoblotting of B Cell Signaling
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B cells cultured under various conditions were harvested and cell lysates were prepared in lysis buffer. The lysate was resolved on a 10% reducing SDS-polyacrylamide gels and transferred to a polyvinylidene difluoride membrane. After blocking with Tris-buffered saline (pH 7.4) containing 5% dried skimmed milk, the membrane was incubated with antibodies, followed by probing with HRP-conjugated anti-rabbit or -mouse antibody (Sigma). The bands were detected by chemiluminescence with ECL detection reagents (Life Technologies). Antibody to Phospho-STAT6 (Tyr641, 56,554), anti-STAT6 (5,397), anti-p-PKA C(4,781), anti-PKA C-α (5,842), anti-p-CREB (9,198), anti-CREB (9,197), PI3 Kinase p110 δ 34,050, anti-p-Akt (Ser473, 4,060), anti-Akt (4,685), anti-p-FoxO1 (Ser256, 9,461), anti-FoxO1 (2,880), anti-p-FoxO3a (Ser253, 13,129), anti-FoxO3a (12,829), anti-PPARγ (2,443) were from Cell Signaling Technology (Danvers, MA, USA). Anti-ubiquitin (PTM-1106) was from PTM BIO (Beijing, China). Anti-β-Actin (66009-1-Ig) was from Proteintech.
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