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17-10531

Sigma-Aldrich

ChIPAb+ LSD1 (KDM1A) Antibody - ChIP Validated Antibody and Primer Set

from rabbit, purified by affinity chromatography

Synonym(s):

Lysine-specific histone demethylase 1A, BRAF35-HDAC complex protein BHC110, Flavin-containing amine oxidase domain-containing protein 2

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.32

biological source

rabbit

Quality Level

clone

polyclonal

purified by

affinity chromatography

species reactivity

rat, human, mouse

manufacturer/tradename

ChIPAb+
Upstate®

technique(s)

ChIP: suitable
immunoprecipitation (IP): suitable
western blot: suitable

NCBI accession no.

UniProt accession no.

shipped in

dry ice

General description

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.
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.

Immunogen

Histidine-tagged recombinant protein corresponding to full length LSD1.

Application

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.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Histones
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.

Packaging

25 assays per set. Recommended use: 2 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).

Quality

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.

Target description

~110 kDa observed. Uniprot describes two isoforms at ~92 kDa and ~95 kDa due to alternative splicing

Physical form

Affinity purified
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

Storage and Stability

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.

Analysis Note

Control
Includes normal rabbit IgG and primers specific for a human prostate-specific antigen (PSA) enhancer region.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage Class Code

10 - Combustible liquids


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Yuhong Lu et al.
Molecular cancer research : MCR, 16(10), 1458-1469 (2018-06-24)
The development of small-molecule tyrosine kinase inhibitors (TKI) specific for epidermal growth factor receptors (EGFR) with activating mutations has led to a new paradigm in the treatment of non-small cell lung cancer (NSCLC) patients. However, most patients eventually develop resistance.
Na Sang et al.
MedComm, 4(3), e269-e269 (2023-05-30)
Lysine-specific histone demethylase 1 (LSD1) is an attractive target for malignancies therapy. Nevertheless, its role in hepatocellular carcinoma (HCC) progression and the potential of its inhibitor in HCC therapy remains unclear. Here, we show that LSD1 overexpression in human HCC
Xin Hu et al.
Oncogene, 38(44), 7017-7034 (2019-08-15)
LSD1 (KDM1A) is a histone demethylase that plays both oncogenic and tumor suppressor roles in breast cancer. However, the exact contexts under which it plays these opposite functions remain largely elusive. By characterizing its role in luminal breast epithelial cells
Lei Tao et al.
Nature communications, 14(1), 7430-7430 (2023-11-17)
Poly (ADP-ribose) polymerase inhibitors (PARPi) are selectively active in ovarian cancer (OC) with homologous recombination (HR) deficiency (HRD) caused by mutations in BRCA1/2 and other DNA repair pathway members. We sought molecular targeted therapy that induce HRD in HR-proficient cells
Nina M Patrick et al.
The Journal of steroid biochemistry and molecular biology, 167, 1-13 (2016-09-21)
Small molecule inhibitors of lysine deacetylases (KDACs) are approved for clinical use in treatment of several diseases. Nuclear receptors, such as the glucocorticoid receptor (GR) use lysine acetyltransferases (KATs or HATs) and KDACs to regulate transcription through acetylation and deacetylation

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