For Western blot analysis, the media were removed from test plates, and cells were lysed in 2× SDS buffer (20% glycerol, 4% sodium dodecyl sulfate, 0.2 M dithiothreitol, 125 mM Tris, pH 6.8), as described.59 (link) Total protein was separated on 4–12% Bis-Tris gels (Life Technologies), transferred to PVDF membranes, and immunoblotted with anti-Pax2 antibody,20 (link) anti-EGFP antibody (Santa Cruz), antiactin (Cell Signaling Tech.), anti-P-Histone H3 (Cell Signaling Tech.), anti-Histone H3 (Cell Signaling Tech.), or anti-Actin (Sigma).
Anti p histone h3
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-p-Histone H3 is a primary antibody that specifically binds to histone H3 phosphorylated at serine 10 or serine 28. This antibody can be used to detect and quantify histone H3 phosphorylation, which is a crucial epigenetic modification associated with various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Vector abc kit, by Vector Laboratories (3 mentions)
Vectastain abc kit, by Vector Laboratories (3 mentions)
3 3 diaminobenzidine dab peroxidase substrate kit, by Vector Laboratories (2 mentions)
Anti p yap, by Cell Signaling Technology (2 mentions)
Anti wisp2, by Cell Signaling Technology (2 mentions)
Anti cleaved caspase 3 antibody, by Cell Signaling Technology (2 mentions)
Anti p erk1 2, by Cell Signaling Technology (2 mentions)
Anti actin, by Merck Group (1 mentions)
Anti egfp antibody, by Santa Cruz Biotechnology (1 mentions)
Anti histone h3, by Cell Signaling Technology (1 mentions)
Anti actin, by Cell Signaling Technology (1 mentions)
Bis tris gel, by Thermo Fisher Scientific (1 mentions)
Sc 56, by Santa Cruz Biotechnology (1 mentions)
Sc 245, by Santa Cruz Biotechnology (1 mentions)
Anti gapdh gtx100118, by GeneTex (1 mentions)
Anti gfp, by Merck Group (1 mentions)
Anti cyclin b1, by Santa Cruz Biotechnology (1 mentions)
Dab chromogen, by Agilent Technologies (1 mentions)
Envision system hrp labelled polymer anti rabbit, by Agilent Technologies (1 mentions)
Anti psmad3, by Thermo Fisher Scientific (1 mentions)
7 protocols using anti p histone h3
1
Western Blot Analysis of Protein Expression
2
Immunoblotting Analysis of Cell Signaling
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Cell lysis and immunoblotting were performed as described [55 (link)]. Proteins were detected using anti-ACC, anti-pSer79-ACC, anti-AMPKα2, anti-pThr172-AMPK, anti-FASN, anti-Cyclin B1 (sc-245, Santa Cruz, Biotechnology, CA, USA), anti-pHistoneH3 (#9701, Cell Signaling), anti-GFP (G6539, Sigma) and anti-α-tubulin. GAPDH and PCNA were detected with anti-GAPDH (GTX100118, GeneTex) and anti-PCNA (sc-56, Santa Cruz) as the loading controls.
3
Immunohistochemical Analysis of Tumor Markers
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Formalin-fixed tumor samples from Day 10 post tumor were deparaffinised and antigen retrieval performed with Citrate buffer. After peroxidase inhibitor incubation and blocking, sections were incubated with either anti-Ki67 (Cell Signaling Technology 12202), anti-pHistone H3 (Cell Signaling Technology 9701 S) or anti-pSMAD3 (Thermo Scientific PA5-110155) antibodies overnight at 4 °C. After washing, sections were incubated with EnVision System HRP Labelled Polymer Anti-Rabbit (Dako k4003), DAB chromogen (Agilent Technologies K346811-2), and counterstained with Mayer’s hematoxylin. Finally, sections were dehydrated, xylene washed as mounted.
4
Macrophage Polarization Assay
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Cell culture media (DMEM high glucose, RPMI 1640), sterile PBS, Trypsin-EDTA and antibiotics (Antibiotic-antimycotic; Penicillin/Streptomycin) were purchased from Life Technology/Invitrogen (Carlsbad, CA, US). Fetal bovine serum (FBS) was from Atlanta Biologicals (Flowery Branch, GA). Primary and secondary antibodies used for immunohistochemistry, immunoblotting and flow cytometry and are as follows: anti-HO-1 (Abcam, Cambridge, MA), anti-β-Actin (Sigma-Aldrich, MO), anti-Notch1 and anti-cleaved Notch1 (Cell Signaling, Beverly, MA; Novus Biologicals; Proteintech), anti-c-myc, anti-P-MAPK-Erk1/2, anti-P-Akt, anti-P-Retinoblastoma (Rb) and anti-P-Histone H3 (Cell Signaling, Beverly, MA), anti-mTFA, anti-P-Elk1/2 and anti-Elk1 (Santa Cruz Biotechnology, Dallas, Texas), anti-cyclin D1 (Calbiochem, Millipore, USA), anti-CD86 and anti-MMR (Biolegend, San Diego, CA). SuperSignal West Femto/Pico Substrate reagents for Western-blot were purchased from Fisher Scientific. Recombinant M-CSF, IL-4 and IFNγ were purchased from Peprotech. LPS (E.coli 0127:B8) was from Sigma. anti-CD86 neutralizing antibodies were purchased from Biolegend. Pegylated catalase (3000 U/mL) and SOD (10 U/mL) were from Sigma and were used 1 hour before CO treatment as previously described [8 (link)].
5
Immunohistochemical Analysis of Tumor Markers
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Primary tumor masses were excised and fixed in 4% paraformaldehyde in PBS overnight. For immunochemistry related studies, sections were deparaffinized, rehydrated with xylene and a descending alcohol gradient, and incubated in 0.3% H2O2. Following antigen retrieval using 10 mM sodium citrate (pH 6.0), sections were incubated with anti-WISP2, anti-p-ERK1/2, anti-p-YAP, anti-p-Histone H3, anti-cleaved caspase-3 antibodies (Cell Signaling Technology, 1:200) using a Vector ABC kit (Vector Laboratories) at room temperature for 1 h. Afterward, the sections were allowed to react with biotin-labeled secondary antibodies for 30 min. Staining was performed using the Vectastain ABC kit and 3,3′-diaminobenzidine (DAB) peroxidase substrate kit (Vector Laboratories, Burlingame, CA, USA).
6
Immunohistochemical Analysis of Tumor Markers
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Primary tumor masses were excised and xed in 4% paraformaldehyde in PBS overnight. For immunochemistry related studies, sections were depara nized, rehydrated with xylene and a descending alcohol gradient, and incubated in 0.3% H 2 O 2 . Following antigen retrieval using 10 mM sodium citrate (pH 6.0), sections were incubated with anti-WISP2, anti-p-ERK1/2, anti-p-YAP, anti-p-Histone H3, anti-cleaved caspase-3 antibodies (Cell Signaling Technology, 1:200) using a Vector ABC kit (Vector Laboratories) at room temperature for 1 h. Afterward, the sections were allowed to react with biotin-labeled secondary antibodies for 30 min. Staining was performed using the Vectastain ABC kit and 3,3′-diaminobenzidine (DAB) peroxidase substrate kit (Vector Laboratories, Burlingame, CA, USA).
7
Immunohistochemical Analysis of Tumor Markers
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Primary tumor masses were excised and fixed in 4% paraformaldehyde in PBS overnight.
For the purpose of immunochemistry related studies, sections were deparaffinized, rehydrated with xylene and a descending alcohol gradient, and incubated in 0.3% H 2 O 2 .
Following antigen retrieval using 10 mM sodium citrate (pH 6.0), sections were incubated with anti-WISP2, anti-p-ERK1/2, anti-p-YAP, anti-p-Histone H3, anti-cleaved caspase 3 (Cell signaling Technology) (1:200) antibodies using a Vector ABC kit (Vector Laboratories), at room temperature for 1 h, and allowed to react with biotin-labeled secondary antibodies for 30 min. Staining was performed using Vectastain ABC kits and 3, 3′-diaminobenzidine (DAB) peroxidase substrate kits (Vector Laboratories, Burlingame, CA, USA).
For the purpose of immunochemistry related studies, sections were deparaffinized, rehydrated with xylene and a descending alcohol gradient, and incubated in 0.3% H 2 O 2 .
Following antigen retrieval using 10 mM sodium citrate (pH 6.0), sections were incubated with anti-WISP2, anti-p-ERK1/2, anti-p-YAP, anti-p-Histone H3, anti-cleaved caspase 3 (Cell signaling Technology) (1:200) antibodies using a Vector ABC kit (Vector Laboratories), at room temperature for 1 h, and allowed to react with biotin-labeled secondary antibodies for 30 min. Staining was performed using Vectastain ABC kits and 3, 3′-diaminobenzidine (DAB) peroxidase substrate kits (Vector Laboratories, Burlingame, CA, USA).
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