Polyclonal antibodies for SAMD1 and L3MBTL3 were made using purified glutathione S-transferase (GST)–fusion proteins as antigen. For SAMD1, the antibody is directed against the SAM domain of human SAMD1 (amino acids 452 to 538). For L3MBTL3, the antibodies are directed against the N terminus (amino acids 3 to 233) or the C terminus (amino acids 778 to 883) of mouse L3MBTL3. Antibodies were made with Eurogentec using the 28-day speedy protocol. Obtained antibodies were affinity-purified. The following commercial antibodies were used: H3K4me3 (Diagenode, C15410003), H3K4me2 (Diagenode, C15410035), H3K27ac (Active Motif, 39133), H3K27me3 (Diagenode, C15410195), KDM1A (Abcam, 17721), tubulin (Millipore, MAB3408), Pou5f1/Oct4 (Santa Cruz Biotechnology, SC-5279), FLAG (Sigma-Aldrich, F3165), hemagglutinin (Roche, 11867423001), H2AUb1 (Cell Signaling Technology, 8240), Lamin B (Santa Cruz Biotechnology, sc-6217), EZH2 (Diagenode, C15410039), L3MBTL2 (Active Motif, 39569), PCGF6 (ProteinTech, 24103-1-AP), RING2 (Abcam, ab101273), RYBP (Sigma-Aldrich, PRS2227), SFMBT1 (Bethyl Laboratories, A303-221A), rabbit immunoglobulin G (IgG) control (Diagenode, C15410206), and H2Av (Drosophila) (Active Motif, 39716). The Sp1 antibody was described previously (42 (link)).
Kdm1a
Manufactured by Abcam
Sourced in United States
KDM1A is a protein involved in the regulation of gene expression. It functions as a histone demethylase, specifically removing methyl groups from histone H3 at lysine 4 (H3K4). This enzymatic activity plays a role in the epigenetic control of gene transcription.
Automatically generated - may contain errors
Lab products found in correlation
Flag tag (flag), by Merck Group (1 mentions)
H3k4me2, by Diagenode (1 mentions)
L3mbtl2, by Active Motif (1 mentions)
H3k27me3, by Diagenode (1 mentions)
H3k4me3, by Diagenode (1 mentions)
H3k27ac, by Active Motif (1 mentions)
Lamin b, by Santa Cruz Biotechnology (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ripa buffer, by Beyotime (1 mentions)
β actin, by Abcam (1 mentions)
Uvchem detection device, by Analytik Jena (1 mentions)
Amersham ecl plus, by Cytiva (1 mentions)
Amersham ecl, by Cytiva (1 mentions)
Hrp conjugated secondary antibody, by OriGene (1 mentions)
Gapdh, by Abcam (1 mentions)
Enhanced chemiluminescence kit, by BestBio (1 mentions)
3 protocols using kdm1a
1
Antibody Generation and Validation for Epigenetic Regulators
2
Western Blot Analysis of Protein Expression
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Total proteins were isolated from cells using RIPA buffer (Beyotime Biotechnology, Shanghai, China), separated by dodecyl sulfate, sodium salt-Polyacrylamide gel electrophoresis (SDS-PAGE, 10%), and transferred onto polyvinylidene didluoride (PVDF) membranes (Millipore, Billerica, MA, USA). The membranes were blocked by 5% skimmed milk at room temperature for 1 h, followed by incubation with primary antibodies (1: 1,000) at 4°C overnight. After washing with a TBST (Tris-buffered saline containing 0.1% Tween-20) buffer three times, the membranes were incubated with HRP-linked secondary antibodies (1:2,000) at room temperature for 1 h and then washed with a TBST buffer for five times. The proteins were visualized by Amersham ECL Plus (Amersham, Arlington Heights, IL, USA) and detected with UVchem Detection Device (Biometra, Gottingen, Germany). Primary antibodies against KDM1A, ZNF346, or β-actin (Abcam, Cambridge, MA, USA) were used in this study.
3
Detecting KDM1A Expression in OCL-LY8 Cells
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Western blotting was used to detect KDM1A protein expression in OCL-LY8 cells. At 48 hours following transfection, total protein was extracted from cell specimens using lysis buffer containing 1% phenylmethanesulfonyl fluoride. Protein concentration was measured by bicinchoninic acid kit. Then, 50 µg protein specimens were separated by 10% SDS-PAGE and electrophoresed onto polyvinylidene difluoride membranes membranes. The membranes were blocked with 5% non-skim milk at room temperature for 2 hours, and incubated with rabbit monoclonal antibodies against KDM1A (1:1,000, Abcam USA, Cambridge, MA, USA) and GAPDH (1:2,000, Abcam USA) at 4°C overnight. HRP-conjugated secondary antibody (1:1,000, OriGene Technologies, Inc., Beijing, People’s Republic of China) was used to incubate the membranes for 30 minutes at room temperature. Finally, signals were detected by the enhanced chemiluminescence kit (BestBio, Shanghai, People’s Republic of China) for 3–5 minutes at room temperature.
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