The following primary rabbit antibodies were purchased from Abcam: anti-H1.2, ab17677; H1.5, ab18208; HMGB2, ab124670; Ki67, ab15580; H3S10p, ab5176; CTCF, ab128873; SMC2, ab10412; RAD21, ab154769; H1x, ab31972. From Cell Signaling: anti-HMGN1, #5692; HMGN2, #9437. From Sigma-Aldrich: anti-H1.4, H7665. From Millipore: anti-H2A, #07-146. The following mouse antibodies were purchased from Abcam: anti-H3, ab24834; H4, ab31830; H2B, ab52584. From Active Motif: anti-H3, #61475. mAb PL2-6 was a gift from M. Monestier (Temple Univ.), see previous use [26 ,27 ]. Secondary antibodies included Invitrogen Alexa 568 goat anti-rabbit IgG, Sigma-Aldrich Atto 488 goat anti-rabbit IgG and Atto 488 goat anti-mouse IgG.
Ab24834
Manufactured by Abcam
Sourced in United States
Ab24834 is a laboratory equipment product. It is designed for general laboratory use.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti rabbit igg atto 488, by Merck Group (1 mentions)
Anti h2a, by Merck Group (1 mentions)
Ab17677, by Abcam (1 mentions)
Ab18208, by Abcam (1 mentions)
Ab31830, by Abcam (1 mentions)
Anti h3, by Active Motif (1 mentions)
Hmgn2, by Cell Signaling Technology (1 mentions)
Atto 488 goat anti mouse igg, by Merck Group (1 mentions)
V5 tag, by Thermo Fisher Scientific (1 mentions)
Gapdh, by Gentex (1 mentions)
Ionic detergent compatibility reagent, by Thermo Fisher Scientific (1 mentions)
Ab32058, by Abcam (1 mentions)
D5671, by Merck Group (1 mentions)
G9545, by Abcam (1 mentions)
Laemmli buffer, by Bio-Rad (1 mentions)
Pierce 660 nm protein assay, by Thermo Fisher Scientific (1 mentions)
Ab2851, by Cell Signaling Technology (1 mentions)
Ab2850, by Abcam (1 mentions)
Ab81371, by Cell Signaling Technology (1 mentions)
A1978, by Abcam (1 mentions)
8 protocols using ab24834
1
Antibody Immunostaining Protocol
2
Immunoblotting Protocol for Protein Detection
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Immunoblotting was performed using standard protocols. Antibodies were diluted in 5% milk powder in TBS-T, and protein detection was carried out with horseradish peroxidase-coupled secondary antibodies (ThermoFisher Scientific, A16110 and A16072) and exposed to X-ray film. The following primary antibodies were used: Olig2 (1:3000; Millipore, AB9610), GAPDH (1:1000; GenTex, GTX627408), p53 (1:500; Cell Signaling, 2524), V5 tag (1:1000; eBioscience, 14679682); histone H3 (1:2000; Abcam, AB24834).
3
Protein Quantification and Immunoblotting
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Cells were collected and directly lysed in Laemmli buffer. Proteins were quantified using Pierce solution supplemented with Ionic Detergent Compatibility Reagent (Thermo Scientific), following manufacturer’s instructions. Equal amounts of cells were loaded for immunoblotting, and sample loading was assessed by Ponceau staining after membrane transfer. Antibodies against SUMO2 (MBL, M114-3), SUMO1 (Abcam, ab32058), Ubc9 (Abcam, ab75854), Dppa2 (Millipore, MAB4356), Dppa4 (R&D Systems, AF3730), Hnrnpc (Abcam, ab133607), Histone H3 (Abcam, ab24834), Smchd1 (Abcam, ab31865), HA-probe (Santa Cruz, SC805), Sae1 (Abcam, ab185949), Uba2 (Abcam, ab185955), Pias1 (Cell Signaling, 3550), Pias2 (Novus Biologicals, NBP2-19819), Pias3 (Santa Cruz, SC46682), Pias4 (Cell Signaling, 4392), Znf451 (Sigma, SAB2108741), Ranbp2 (Santa Cruz, SC74518), Senp1 (Santa Cruz, SC271360), Senp2 (Santa Cruz, SC67075), Senp3 (Cell Signaling, 5591), Senp5 (Abcam, ab47631), Senp6 (Thermo Fisher Scientific, PA5-69704), and Senp7 (Thermo Fisher Scientific, PA5-36089), were used between 1/1000 and 1/500 concentration according to standard protocols and supplier’s recommendations.
4
Cell Line Culture and Protein Analysis
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The human cervical cancer cell line HeLa and human lung cancer cell line A549 were cultured in Dulbecco's Modified Eagle's Medium (Sigma-Aldrich D5671) supplemented with 10% FBS and 1% penicillin/streptomycin at 37°C and 5% CO2. Anti-GAPDH antibody (Sigma-Aldrich G9545) and anti-histone H3 antibody (Abcam ab24834) were used for western blot analysis.
5
Protein Expression and Quantification
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Cells were collected and directly lysed in Laemmli buffer (Bio-Rad #161–0747). Proteins were quantified using Pierce 660 nm Protein Assay (ThermoFisher Scientific #22662) according to manufacturer’s instructions. Equal amounts of proteins were loaded on gels and good equilibration of the different samples was assessed by Ponceau staining after membrane transfer. Antibodies against SUMO1 (1:1000, Abcam #ab32058), Actin (1:4000, Sigma #A1978), SUMO2/3 (1:1000, Abcam #ab81371), Gapdh (1:1000, Cell Signaling #2118), Dnmt3a (1:1000, Abcam #ab2850), Histone H3 (1:5000, Abcam #ab24834), Dnmt3b (1:1000, Abcam #ab2851), Tet2 (1:1000, Cell Signaling #36449S) were used according to standard protocols and suppliers’ recommendations.
6
Detecting Histone H3 Citrullination
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Antibodies for detecting citrullinated histone H3 (R2 citrullination, Ab176843 and R2-8-17 citrullination, Ab5103), total histone H3 (Ab24834), NCF1, NCF2, and NCF4 were from Abcam (Cambridge, MA). Antibodies for detecting PAD4 (MABE254) and citrullination (F95) were from EMD Millipore (Billerica, MA). Antibodies for PAD IP were developed as previously described40 (link). Recombinant human PAD2 and recombinant human PAD4 were generated in-house. Recombinant human histone H3, pan PAD inhibitor BB-Cl-amidine, and PAD4 inhibitor GSK484 were from Cayman Chemicals (Ann Arbor, MI). The cell fraction isolation kit was from Cell Signaling (Danvers, MA). Protein G beads, DPI, ionomycin, PMA, Hoechst 33342, HRP-labeled anti-mouse immunoglobulin G (IgG), and anti-rabbit IgG secondary antibodies were from Invitrogen (Carlsbad, CA).
7
Antibody Characterization for EBOV Studies
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The antibodies used in this study include monoclonal anti-polyhistidine−alkaline phosphatase antibody (Milliposre Sigma #A5588, St. Louis, MO, USA), anti-EGFP monoclonal antibody (ThermoFisher Scientific #MA1-952, Grand Island, NY, USA), anti-HA tag antibody (Abcam #ab18181, Cambridge, MA, USA), anti-EBOV VP24 polycolonal antibody (IBT Bioserveices #0301-047, Rockville, MD, USA), anti-EBOV VP30 polycolonal antibody (IBT Bioserveices #0301-048, Rockville, MD, USA), anti-EBOV VP40 polycolonal antibody (IBT Bioserveices #0301-010, Rockville, MD, USA), anti-GAPDH monoclonal antibody (6C5) (ThermoFisher Scientific #AM4300, Grand Island, NY, USA), anti-alpha 1 sodium potassium ATPase antibody (Abcam #ab7671, Cambridge, MA, USA), anti-LAMP2 antibody (Abcam #ab13524, Cambridge, MA, USA), and anti-histone H3 antibody (Abcam #ab24834, Cambridge, MA, USA).
8
Protein Extraction and Immunoblotting Protocol
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All cell lysis was performed with RIPA buffer (Cell Signaling Technology) with added cOmplete protease inhibitor cocktail (25× dilution; Roche), 1 mmol/L phenylmethylsulfonyl fluoride (Sigma-Aldrich), and phosphatase inhibitor cocktails 2 and 3 (100× dilution; Sigma-Aldrich). Total protein extracted was quantified by the Bradford assay with the Bio-Rad protein dye reagent (Bio-Rad). Quantified protein samples, denatured by heating, were resolved by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to a polyvinylidene difluoride membrane (Merck Millipore). Primary antibodies used for immunoblotting against various targets were listed as follows: PTK7 (1:1000, AF4499; R&D Systems), SOX9 (1:1000, AB553; Merck Millipore), Smad2 (1:1000, 5339; Cell Signaling Technology), pSmad2 (1:1000, 3108; Cell Signaling Technology), SLUG (1:1000, MA5-26385; Thermo Scientific), ZEB1 (1:500, ab203829; Abcam), β-catenin (1:1000, 9587; Cell Signaling Technology), α-tubulin (1:1000, T9026; Sigma-Aldrich), histone H3 (1:2000, ab24834; Abcam), and β-actin (1:10,000, A5316; Sigma-Aldrich).
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