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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, M, R | ChIP, WB, IP |
| Description | |
|---|---|
| Catalogue Number | 17-10531 |
| Trade Name |
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| Description | ChIPAb+ LSD1 (KDM1A) Antibody - ChIP Validated Antibody and Primer Set |
| Overview | All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction. The ChIPAb+ LSD1 (KDM1A) set includes a LSD1 antibody, a Normal rabbit IgG and positive control primers which amplify a 85bp human prostate-specific antigen enhancer region. The LSD1 antibody and negative control IgG are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of LSD1 associated chromatin. |
| Alternate Names |
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| Background Information | LSD1 (lysine specific demethylase 1), also known as BHC110, is a histone demethylase that specifically demethylates Lys4 of histone H3, a specific tag for epigenetic transcriptional activation, thereby acting as a corepressor. LSD1 acts by oxidizing the substrate by FAD to generate the corresponding imine that is subsequently hydrolyzed. LSD1 demethylates both mono- and tri-methylted Lys4 of histone H3, and may play a role in the repression of neuronal genes. Alone, it is unable to demethylate H3 Lys4 on nucleosomes and requires the presence of RCOR1/CoREST to achieve such activity. LSD1 may also demethylate Lys9 of histone H3, a specific tag for epigenetic transcriptional repression, thereby leading to derepression of androgen receptor target genes. |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
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| Presentation | Anti-LSD1 (Rabbit Polyclonal), Part No. CS207350. One vial containing 50 µg affinity purified rabbit polyclonal in 72 µL buffer containing 0.1M Tris-Glycine (pH 7.4), 150 mM NaCl and 0.05% sodium azide, before the addition of glycerol to 30%. Final concentration of antibody 0.7 mg/mL. Store at -20°C. Normal rabbit IgG, Part No. PP64B. One vial containing 125 µg Rabbit IgG in 125 µL storage buffer containing 0.05% sodium azide. Store at -20°C. ChIP Primers, PSA. Part No. CS207340. One vial containing 75 μL of 5 μM of each primer specific for human prostate-specific antigen (PSA) enhancer region (chr19:51354179-51354263, hg19 build). Store at -20°C. FOR: GCC TGG ATC TGA GAG AGA TAT CAT C REV: ACA CCT TTT TTT TTC TGG ATT GTT G |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Use ChIPAb+ LSD1 (KDM1A) ChIP Validated mouse monoclonal antibody for the detection of LSD1 (Lysine-specific histone demethylase 1A) also known as BRAF35-HDAC complex protein BHC110. |
| Key Applications |
|
| Application Notes | Chromatin Immunoprecipitation: Sonicated chromatin prepared from hormone starved (72 hrs), then androgen agonist induced (1 µM CI-4AS-1 overnight) LNCaP cells (~1e5 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Rabbit IgG (Part No. PP64B), or 2 µg Anti-LSD1 (Part No.CS207350) and the Magna ChIP™ HiSens (Cat. # 17-10460). Successful immunoprecipitation of LSD1 associated DNA fragments was verified by qPCR using ChIP Primers, PSA (Part No. CS207340) as a positive locus, and ChIP Primers, β-globin (Part No. CS200591) as a negative locus. Please refer to the Magna ChIP™ HiSens (Cat. # 17-10460) or EZ-Magna ChIP™ HiSens(Cat. # 17-10461) protocol for experimental details. Chromatin Immunoprecipitation: Sonicated chromatin prepared from hormone starved (72 hrs), then androgen agonist induced (1 µM CI-4AS-1 overnight) LNCaP cells (1e5 and 1e6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Rabbit IgG (Part No. PP64B), or 2 µg Anti-LSD1 (Part No. CS207350) and the Magna ChIP™ HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of LSD1 associated DNA fragments was verified by qPCR using ChIP Primers, PSA (Part No. CS207340). Please refer to the Magna ChIP™ HiSens (Cat. # 17-10460) or EZ-MagnaChIP™ HiSens (Cat. # 17-10461) protocol for experimental details Western Blotting Analysis: HeLa nuclear extract was probed with Anti-LSD1 (0.05 μg/mL). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. |
| Biological Information | |
|---|---|
| Immunogen | Histidine-tagged recombinant protein corresponding to full length LSD1. |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Species Reactivity |
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| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Purification Method | Affinity Purfied |
| UniProt Number |
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| Molecular Weight | ~110 kDa observed. Uniprot describes two isoforms at ~92 kDa and ~95 kDa due to alternative splicing |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Chromatin Immunoprecipitation: Sonicated chromatin prepared from hormone starved (72 hrs), then androgen agonist induced (1 µM CI-4AS-1 overnight) LNCaP cells (~1e5 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Rabbit IgG (Part No. PP64B), or 2 µg Anti-LSD1 (Part No.CS207350) and the Magna ChIP™ HiSens Chromatin IP Kit (Cat. # 17-10460). Successful immunoprecipitation of LSD1 associated DNA fragments was verified by qPCR using ChIP Primers, PSA (Part No. CS207340) (Figure 1). Please refer to the MagnaChIP™ HiSens (Cat. # 17-10460) or EZ-MagnaChIP™ HiSens (Cat. # 17-10461) protocol for experimental details. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. |
| Packaging Information | |
|---|---|
| Material Size | 25 assays |
| Material Package | 25 assays per set. Recommended use: 2 μg of antibody per chromatin immunoprecipitation (dependent upon biological context). |