Immunoblotting was performed with the following antibodies: Bax (D2E11, #5023), Bcl-xL (#2762), Caspase-3 (#9662), Caspase-9 (#9502), p-Cdc2 (Tyr15) (#9111), Cdc2 (POH1, #9116), p-CDK2 (Thr160) (#2561), p-Chk2 (Thr68) (C13C1, #2197), Chk2 (#2662), c-Raf (#9422), CDK2 (78B2, #2546), Cyclin D1 (#2922), Cyclin E1 (D7T3U, #20808), DUSP4 (D9A5, #5149), DUSP6 (#39441), p-Erk1/2 (Thr202/Tyr204) (D13.14.4E, #4370), Erk1/2 (137F5, #4695), Mcl-1 (D5V5L, #39224), MDM2 (D1V2Z, #86934), p-MEK1/2 (Ser217/221) (41G9, #9154), MEK1/2 (47E6, #9126), p21 (12D1, #2947), p-p53 (Ser15) (16G8, #9286), p53 (7F5, #2527), PARP (46D11, #9532), PUMA (D30C10, #12450), p-Rb (Ser789) (#9307), (all from Cell Signaling Technology (Boston, MA, USA)), Actin (C-2, #sc-8432 AF790, Santa Cruz Biotechnology), and β-actin antibody (AC-15, #A5441, Sigma-Aldrich). Immunoblotting signals were visualized using an Odyssey IR imaging system (Li-Cor Biosciences, Lincoln, NE, USA). Image Studio software v4.0 was used for analysis of the bands.
β actin antibody ac 15
Manufactured by Merck Group
Sourced in United States
The β-actin antibody (AC-15) is a laboratory reagent used in various research applications. It is a mouse monoclonal antibody that specifically binds to the β-actin protein, a ubiquitous and highly expressed cytoskeletal protein found in all eukaryotic cells. The antibody can be used to detect and quantify the expression levels of β-actin in samples, such as cell lysates or tissue extracts, using techniques like Western blotting, immunocytochemistry, or immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Caspase 3, by Cell Signaling Technology (1 mentions)
C raf, by Cell Signaling Technology (1 mentions)
Actin c 2, by Santa Cruz Biotechnology (1 mentions)
Odyssey ir imaging system, by LI COR (1 mentions)
Bcl xl, by Cell Signaling Technology (1 mentions)
Mcl 1, by Cell Signaling Technology (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
Ago2 antibody, by Abcam (1 mentions)
Lps l2630, by Merck Group (1 mentions)
Pcdna3 flag jnk2a2, by Addgene (1 mentions)
Adenosine triphosphate (atp), by Enzo Life Sciences (1 mentions)
Actinomycin d, by Merck Group (1 mentions)
Anti mouse igg hrp, by GE Healthcare (1 mentions)
Alexa fluor 488 donkey anti goat igg, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
C myc 9e10, by Santa Cruz Biotechnology (1 mentions)
Alexa fluor 594 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 647 donkey anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions)
4 protocols using β actin antibody ac 15
1
Immunoblotting of Cell Signaling Proteins
2
miR-221-5p Regulation of JNK2 Signaling
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LPS (L2630), β-actin antibody (AC-15), and actinomycin D were from Sigma-Aldrich. ATP was from ENZO Life Sciences. JNK2 antibody was from Cell Signaling Technology (Cat # 4672). AGO2 antibody was from abcam (ab186733). Mouse miRIDIAN miR-221-5p mimic, miRIDIAN miRNA mimic negative control, miRIDIAN mouse miR-221-5p hairpin inhibitor, and miRIDIAN miNA hairpin inhibitor negative control were from Horizon Discovery. Ad/JNK2 was generated by ViraQuest, Inc. by cloning pCDNA3 Flag Jnk2a2 (Addgene, Item ID 13755) into the pVQAd CMV K-NpA vector. Ad/null was from ViraQuest, Inc. miR-221-5p inhibitor and inhibitor control, and miR-221-5p mimic and mimic control were from Horizon Discovery.
3
Investigating Phosphorylation Signaling Pathways
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Antibodies against phospho-SHIP1 (Tyr1020), phospho-SHP-1 (Tyr564) (D11G5), phospho-Src family (Tyr416) (D49G4), phospho-p38 MAPK (Thr180/Tyr182) (3D7), phospho-(Ser) PKC substrate, phospho-p40phox (Thr154), and Rac1/2/3 antibodies were purchased from Cell Signaling Technology. Phospho-ERK (E-4), p38α/β MAPK (H-147), p47phox (H-195), p22phox (C-17), Rab5 (D-11), Rab7 (H-50), calnexin (H-70), c-Myc (9E10), and COX-2 (C-20) antibodies were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). The iNOS/NOS type II (54/iNOS) antibody was purchased from BD Biosciences, and the β-actin (AC-15) antibody was purchased from Sigma-Aldrich. Secondary antibodies for Western blotting, ECL™ anti-rabbit, and anti-mouse IgG HRP were obtained from GE Healthcare. For immunofluorescence and confocal laser microscopy, Alexa Fluor®488 goat anti-mouse IgG, Alexa Fluor®488 goat anti-rabbit IgG, Alexa Fluor®488 donkey anti-goat IgG, Alexa Fluor®594 goat anti-rabbit IgG, and Alexa Fluor®647 donkey anti-mouse IgG were obtained from Invitrogen. Phorbol 12-myristate 13-acetate (PMA) and pure lipopolysaccharide (LPS) from E. coli O111:B4 (L3024) were purchased from Sigma-Aldrich. The inhibitors, bisindolylmaleimide I (BIM-1), SB203580, and LY294992 were obtained from Calbiochem (San Diego, CA, USA).
4
Quantifying AhR Expression in THP-1 Macrophages
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The expression of AHR in the nuclei and the cytoplasm of THP‐1 macrophages was assessed as follows. The cells were resolved using NE‐PER Nuclear and Cytoplasmic Extraction Reagents (Life Technologies) with 1% Halt Protease and Phosphatase Inhibitor Cocktail (Life Technologies) according to the manufacturer's instructions.
Protein concentrations were determined using Pierce BCA Protein Assay Kits (Life Technologies). Protein (10 μg) was resolved by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis at 200 V for 30 minutes and transferred to PVDF membranes using X cell II Blot module (Life Technologies). Proteins were detected using anti‐AHR antibody (#83200, AhR [D5S6H]), rabbit mAb, (Cell Signal Technology Japan, Tokyo, Japan), mouse monoclonal anti‐Lamin B1 antibody (sc‐377000; Santa Cruz Biotechnology Inc., Dallas, TX, USA), a mouse monoclonal anti‐β‐actin (AC‐15) antibody (A5441; Sigma‐Aldrich) and the specific secondary antibodies, donkey cy3‐conjugated affinity pure anti rabbit IgG (H + L) antibody (Jackson ImmmunoResearch Laboratories Inc., West Grove, PA, USA) for AHR, goat anti‐mouse IgG (H&L) and Alexa Fluor 647 (ab150119; Abcam) for Lamin B1 and β‐actin, respectively. Fluorescent images were acquired using an ImageQuant LAS‐4000 lumino‐image analyzer (GE Healthcare UK Ltd., Little Chalfont, Buckinghamshire, UK).
Protein concentrations were determined using Pierce BCA Protein Assay Kits (Life Technologies). Protein (10 μg) was resolved by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis at 200 V for 30 minutes and transferred to PVDF membranes using X cell II Blot module (Life Technologies). Proteins were detected using anti‐AHR antibody (#83200, AhR [D5S6H]), rabbit mAb, (Cell Signal Technology Japan, Tokyo, Japan), mouse monoclonal anti‐Lamin B1 antibody (sc‐377000; Santa Cruz Biotechnology Inc., Dallas, TX, USA), a mouse monoclonal anti‐β‐actin (AC‐15) antibody (A5441; Sigma‐Aldrich) and the specific secondary antibodies, donkey cy3‐conjugated affinity pure anti rabbit IgG (H + L) antibody (Jackson ImmmunoResearch Laboratories Inc., West Grove, PA, USA) for AHR, goat anti‐mouse IgG (H&L) and Alexa Fluor 647 (ab150119; Abcam) for Lamin B1 and β‐actin, respectively. Fluorescent images were acquired using an ImageQuant LAS‐4000 lumino‐image analyzer (GE Healthcare UK Ltd., Little Chalfont, Buckinghamshire, UK).
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