Cycloheximide and MG132 were purchased from Cayman Chemical (Ann Arbor, MI, USA). Lithium chloride was obtained from Thermo Fisher Scientific (Waltham, MA, USA). S-protein agarose beads were obtained from Merck (Rahway, NJ, USA). MYC antibody-conjugated agarose beads were from Thermo Fisher Scientific (Waltham, MA, USA). The following antibodies were used: anti-FLAG (Proteintech; San Diego, CA, USA), anti-MYC (Proteintech; San Diego, CA, USA), anti-OTUD7B (Proteintech; San Diego, CA, USA), anti-cyclophilin B (Invitrogen; Waltham, MA, USA), anti-HSP90 (Santa Cruz Biotechnology; Santa Cruz, CA, USA), anti-HA (Santa Cruz Biotechnology; Santa Cruz, CA, USA), anti-V5 (Proteintech; San Diego, CA, USA), anti-α-tubulin (Proteintech; San Diego, CA, USA), and anti-lamin B1 (Santa Cruz Biotechnology; Santa Cruz, CA, USA).
Anti lamin b1
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-Lamin B1 is a laboratory reagent used for the detection and analysis of Lamin B1 protein. Lamin B1 is a structural protein found in the cell nucleus and plays a crucial role in maintaining the structural integrity of the nuclear envelope. The Anti-Lamin B1 product is designed to specifically bind to and detect the Lamin B1 protein, enabling researchers to study its expression, localization, and potential alterations in various biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Anti lamin a c, by Santa Cruz Biotechnology (5 mentions)
Anti β actin, by Santa Cruz Biotechnology (5 mentions)
Anti erk1 2, by Cell Signaling Technology (4 mentions)
Anti akt, by Cell Signaling Technology (3 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (3 mentions)
Anti gfp, by Santa Cruz Biotechnology (3 mentions)
Anti iκbα, by Cell Signaling Technology (3 mentions)
β actin, by Merck Group (3 mentions)
Anti p akt, by Cell Signaling Technology (2 mentions)
Anti foxo1, by Cell Signaling Technology (2 mentions)
Anti pi3k, by Cell Signaling Technology (2 mentions)
Anti phospho erk1 2, by Cell Signaling Technology (2 mentions)
Anti p stat3, by Cell Signaling Technology (2 mentions)
Anti gapdh, by Proteintech (2 mentions)
Anti gapdh, by Cell Signaling Technology (2 mentions)
Anti nfat2, by Santa Cruz Biotechnology (2 mentions)
Hrp conjugated secondary abs, by BD (2 mentions)
Anti β actin, by Merck Group (2 mentions)
Anti phospho erk, by Cell Signaling Technology (2 mentions)
Anti gapdh, by Santa Cruz Biotechnology (2 mentions)
46 protocols using anti lamin b1
1
Investigating MYC-interacting proteins using affinity purification
2
Immunocytochemistry of Fibroblast Cells
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For immunocytochemistry, fibroblasts were grown directly on coverslips. Cells were fixed in 100 % methanol at -20 °C for 10 min. Subsequently, cells were further processed for immunohistochemistry as previously described [45 (link)]. The following primary antibodies were used: anti-progerin S9 (1 µg mL-1) [47 (link)], anti-lamin A (Abcam, 1/500), anti-lamin B1 (Santa Cruz Biotechnology, 1/75), anti-γH2AX (Millipore, 1/200), anti-53BP1 (A300-272A, Bethyl, 1/1000), and anti-Rad51 (Novusbio, 1/300). The secondary antibodies were affinity-purified Alexa Fluor 488 goat or donkey IgG antibodies (Molecular Probes) and Cy3-conjugated IgG antibodies (Jackson ImmunoResearch). DAPI in Vectashield mounting medium (Vector Inc.) was used to counterstain the samples. Images were acquired by using an Axioplan fluorescence microscope (Carl Zeiss).
3
Antibody Characterization in Cell Lines
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The following antibodies were used: mouse monoclonal anti-PARP-1, mouse monoclonal anti-BrdU (#347580), mouse monoclonal anti-p21 (#556431), mouse monoclonal anti-p27 (#610241), rabbit polyclonal anti-PAR (#551813,; BD Bioscience), mouse monoclonal anti-α-tubulin (#T5168, Sigma-Aldrich), mouse monoclonal antibodies for Myc (#G019), HA (#G036), Flag (#G191), and GFP (#G096, Abm), rabbit polyclonal anti-MAP2 (#MAB3418), anti-Olig2 (#ab9610), anti-SOX2 (#ab5603, Millipore), anti-GFAP (#ab7260), anti-Ki67 (#ab15580), anti-Tbr2 (#ab183991) and mouse monoclonal anti-Nestin (#ab22035, abcam), goat polyclonal anti-DCX (#sc-8066), anti-NeuroD (#sc-1084), rabbit polyclonal anti-ERK (#sc-93), mouse monoclonal anti-pERK (#sc-7383), anti-LaminB1 (#sc-56145, Santa Cruz), rabbit polyclonal anti-Akt (#4060), rabbit polyclonal anti-pAkt (#4061), rabbit polyclonal anti-PI3K (#4228), anti-pPI3K (#4257), rabbit polyclonal anti-FOXO1 (#2880), rabbit polyclonal anti-pFOXO1 antibody (#9461, Cell Signaling).
4
Probing Cell Signaling Pathways
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Primary antibodies used in this study were anti-beta actin (Abcam, ab8224), anti-ERK1/2 (Cell Signaling, 9102), anti-phospho ERK1/2 (Cell Signaling, 9101), anti-lamin A/C (Santa Cruz, sc-7292), anti-lamin B1 (Santa Cruz, sc-30264), anti-S6 kinase (Cell Signaling, 2317), anti-phospho S6 kinase (Cell Signaling, 2211) and anti-LAP1 (Atlas antibodies, HPA050546).
5
Synthesis and Characterization of ZYZ-803
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ZYZ-803 was synthesized by the reaction of 2-amino-3-propynylsulfanyl-propionic acid with cinnamyl alcohol and purified as described before [2 ]. WP1066 and KN93 were purchased from Medchemexpress LLC (Monmouth Junction, NJ, USA). PAG and L-NAME were purchased from Sigma-Aldrich (St. Louis, MO, USA). The primary antibodies used were as follows: anti-STAT3 and anti-GAPDH were purchased from Proteintech (Wuhan, Hubei, China); anti-p-STAT3, anti-p-CAMKII and anti-CAMKII (pan) were purchased from Cell Signaling Technology (Beverly, MA, USA); anti-PCNA and anti-LaminB1 were from Santa Cruz Biotechnology (Santa Cruz, CA, USA); and anti-cyclin D1 was from Bioss (Beijing, China). Horseradish peroxidase (HRP)-conjugated secondary antibodies were from Jackson Laboratories (West Grove, PA, USA). The EdU Assay Kit was purchased from Beyotime Institute of Biotechnology (Shanghai, China).
6
Antibody Panel for DNA Damage Response
7
Subcellular Fractionation and Immunoblotting
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Cultured cells were harvested after washing with ice-cold PBS and lysed in extraction buffer (50 mM Trsi-HCl, pH 8.0, 150 mM NaCl, 1 mM EDTA, 0.5% Nonidet P-40, 0.01% protease inhibitor mixture). Cells were fractionated using Nuclear and Cytoplasmic Extraction reagents (Pierce). Cytoplasmic and nuclear extracts were subjected to SDS-PAGE and transferred onto nitrocellulose. Membranes were blocked for 1h with skim milk in Tris-buffered saline containing 0.1% Tween 20% and incubated overnight at 4°C with anti-NFAT2 (0.2 μg/ml), anti-lamin B1 (0.1 μg/ml; Santa Cruz) and anti-β-actin (2.2 μg/ml; Sigma). Membranes were washed, incubated for 1h with HRP-conjugated secondary Abs (BD Biosciences), and developed using chemiluminescent substrates.
8
Analyzing Anti-SMOC2 Protein Interactions
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Anti-SMOC2 was purchased from OriGene (catalog number: TA351730). Anti-β-actin antibody was purchased from Abcam (Cambridge, UK, catalog number: ab6276). The anti-caspase-3 (catalog number: #9662), anti-cleaved caspase-3 (catalog number: #9661), anti-beta-catenin (catalog number: #9562), anti-ERK (catalog number: #4695), anti-AKT (catalog number: #4691), anti-phospho-ERK (catalog number: #4370), and anti-phospho-AKT (catalog number: #4060), anti-cleaved PARP (catalog number: #5625), anti-BAX (catalog number: #5023), and anti-BIM (catalog number: #2933) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-LaminB1 (catalog number: sc-365214), anti-mouse IgG-HRP (catalog number: sc-2031) and anti-rabbit IgG-HRP (catalog number: sc-2030) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Geneticin was purchased from Sigma Aldrich (St Louis, MO, USA). Recombinant human SMOC2 protein was purchased from R&D systems (Minneapolis, MN, USA, catalog number: 5140-SM-050).
9
Antibody Profiling for Cell Stress Signaling
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The following antibodies were used in the study: anti-GRP78 (sc-1050, Santa Cruz, CA, USA), anti-p-PERK Thr981 (sc-32577, Santa Cruz), anti-eIf2α (sc-133132, Santa Cruz), anti-CRT (sc-166837, Santa Cruz), anti-ERp57 (sc-28823, Santa Cruz), anti-Ub (sc-8017, Santa Cruz), anti-p-Akt1/2/3 (sc-7985, Santa Cruz), anti-GAPDH (sc-32233, Santa Cruz), anti-GFP (sc-8334, Santa Cruz), anti-Lamin B1 (sc-374015, Santa Cruz), anti-CaM (sc-137079, Santa Cruz), anti-CaMKIIγ (sc-1541, Santa Cruz), CREB-1 (sc-186, Santa Cruz), anti-p-CaMKII (ab182647, abcam, Waltham, MA, USA), anti-Bcl-2 (610539, BD Biosciences, Franklin Lakes, NJ, USA), anti-cleaved-caspase 3 (9664, Cell Signaling Technology, Danvers, MA, USA), anti-p-CREB Ser133 (9198, Cell Signaling Technology), anti-p-eIF2α Ser51 (9721, Cell Signaling Technology), and anti-PARP (9542, Cell Signaling Technology). All secondary antibodies (HRP-conjugated anti-rabbit, anti-mouse, and anti-goat) were purchased from Sigma (Sigma-Aldrich Inc., St. Louis, MO, USA). All the drug formulations were purchased from Sigma (Sigma-Aldrich Inc.).
10
Mechanistic Insights into PIWIL2-p65 Pathway Regulation
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CDS encoding PIWIL2 and p65 was synthesized and inserted into pcDNA3.1 and pEGFP, respectively. shRNA against PIWIL2 and p65 were synthesized by TSINGKE Biological Technology company (Beijing, China) and the target sequences of these shRNA were as follows:
Human PIWIL2 shRNA (shPIWIL2-1): 5′-CGG ATT GAG GAG AAA CGT AAA CTC-3′
Human PIWIL2 shRNA (shPIWIL2-2): 5′-CTA TGA GAT TCC TCA ACT ACA GAA G-3′
Human p65 shRNA (shp65): 5′ -CGG ATT GAG GAG AAA CGT AAA CTC-3′
Anti-PIWIL2, anti-p62, anti-Beclin-1, and anti-LaminB1 were purchased from Santa Cruz (USA). Anti-IKKα, anti-IKKβ, anti-p-IKKα/β(Ser176/180), anti-IκBα, anti-p-IκBα(Ser32/36), anti-p65, anti-p-mTOR(Ser2448), anti-p-ULK1(Ser555), anti-p-4E-BP1(Thr37/46), anti-p-P70S6(Thr389), and anti-TSC1 were purchased from Cell signaling technology (USA). Anti-LC3B was purchased from Novus Biologicals (USA). Anti-mTOR and anti-Bcl-2 were purchased from Proteintech Group (USA). Anti-GAPDH was purchased from Epitomics (USA).
Human PIWIL2 shRNA (shPIWIL2-1): 5′-CGG ATT GAG GAG AAA CGT AAA CTC-3′
Human PIWIL2 shRNA (shPIWIL2-2): 5′-CTA TGA GAT TCC TCA ACT ACA GAA G-3′
Human p65 shRNA (shp65): 5′ -CGG ATT GAG GAG AAA CGT AAA CTC-3′
Anti-PIWIL2, anti-p62, anti-Beclin-1, and anti-LaminB1 were purchased from Santa Cruz (USA). Anti-IKKα, anti-IKKβ, anti-p-IKKα/β(Ser176/180), anti-IκBα, anti-p-IκBα(Ser32/36), anti-p65, anti-p-mTOR(Ser2448), anti-p-ULK1(Ser555), anti-p-4E-BP1(Thr37/46), anti-p-P70S6(Thr389), and anti-TSC1 were purchased from Cell signaling technology (USA). Anti-LC3B was purchased from Novus Biologicals (USA). Anti-mTOR and anti-Bcl-2 were purchased from Proteintech Group (USA). Anti-GAPDH was purchased from Epitomics (USA).
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