LSD1 Antibody - BSA Free

Novus Biologicals | Catalog # NBP3-18684

Novus Biologicals
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Key Product Details

Species Reactivity

Human

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Chromatin Immunoprecipitation (ChIP), Chromatin Immunoprecipitation Sequencing

Label

Unconjugated

Antibody Source

Polyclonal Rabbit

Format

BSA Free
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Product Specifications

Immunogen

Polyclonal antibody raised in rabbit against human LSD1 (Lysine-specific demethylase 1), using a KLH-conjugated synthetic peptide from the inner part of the protein.

Clonality

Polyclonal

Host

Rabbit

Scientific Data Images for LSD1 Antibody - BSA Free

Western Blot: LSD1 Antibody [NBP3-18684]

Western Blot: LSD1 Antibody [NBP3-18684]

Western Blot: LSD1 Antibody [NBP3-18684] - Figure 5. Western blot analysis using the antibody directed against LSD1 Whole cell extracts (40 ug) from HeLa cells transfected with LSD1 siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the antibody against LSD1 diluted 1:5,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
Immunocytochemistry/ Immunofluorescence: LSD1 Antibody [NBP3-18684]

Immunocytochemistry/ Immunofluorescence: LSD1 Antibody [NBP3-18684]

Immunocytochemistry/Immunofluorescence: LSD1 Antibody [NBP3-18684] - Figure 6. Immunofluorescence using the antibody directed against LSD1 HeLa cells were stained with the antibody against LSD1 and with DAPI. Cells were fixed with 4% formaldehyde for 10 and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the LSD1 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
Chromatin Immunoprecipitation Sequencing: LSD1 Antibody [NBP3-18684] - Figure 2. ChIP-seq results obtained with the antibody directed against LSD1 ChIP was performed on sheared chromatin from 4,000,000 K562 cells using 1 ug of the antibody against LSD1 as described above. The IPd DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturers instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 600 kb region of the X-chromosome (figure 2A and B) and in three regions surrounding the MYT1, RBM19 and TGFBR3 positive control genes, respectively (figure 2C, D and E). The position of the amplicon used for ChIP-qPCR is indicated by an arrow.
Western Blot: LSD1 Antibody [NBP3-18684]

Western Blot: LSD1 Antibody [NBP3-18684]

Western Blot: LSD1 Antibody [NBP3-18684] - Figure 4. Western blot analysis using the antibody directed against LSD1 Western blot was performed using nuclear extracts from HeLa cells (40 ug) and the antibody against LSD1 diluted 1:4,000 in TBS- Tween containing 5% skimmed milk. The molecular weight marker (in kDa) is shown on the left. The location of the protein of interest is indicated on the right.
Chromatin Immunoprecipitation Sequencing: LSD1 Antibody [NBP3-18684] - Figure 1. ChIP results obtained with the antibody directed against LSD1 ChIP was performed with the antibody against LSD1 on sheared chromatin from 4,000,000 K562 cells using the "iDeal ChIP-seq" kit.. An antibody titration consisting of 1, 2, 5 and 10 ug per ChIP experiment was analysed. IgG (2 ug/IP) was used as negative IP control. QPCR was performed with primers for specific regions in the MYT1, RBM19, and TGFBR3 genes, used as positive controls, and for the MYOD1 gene, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
ELISA: LSD1 Antibody [NBP3-18684]

ELISA: LSD1 Antibody [NBP3-18684]

ELISA: LSD1 Antibody [NBP3-18684] - Figure 3. Determination of the antibody titer To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against LSD1 in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:176,000.

Applications for LSD1 Antibody - BSA Free

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

1 ug/ChIP

Chromatin Immunoprecipitation Sequencing

1 ug/ChIP

ELISA

1:1000-1:10000

Immunocytochemistry/ Immunofluorescence

1:200

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Paraffin

1:200-1:500

Western Blot

1:4000
Application Notes
Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 ug per IP.

Formulation, Preparation, and Storage

Purification

Affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide and 0.05% ProClin 300

Concentration

2.2 mg/ml

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.

Background: Lysine (K)-specific Demethylase 1A/KDM1A/LSD1

LSD1 a recently identified, first known lysine-specific histone demethylase, is an 886 amino acid nuclear protein belonging to flavin monoamine oxidase family. It contains a SWIRM domain, a FAD-binding motif and an amine oxidase domain. This protein is ubiquitously expressed and is a component of several histone deacetylase complexes. LSD1 acts as a component of the CoREST and other transcriptional co-repressor complexes and also plays an important role in silencing neuronal-specific genes in non-neuronal cells. It is also known to demethylate Lys-4 of histone H3, a specific tag for epigenetic transcriptional activation. Reports suggest that it plays an important role in stimulating androgen-receptor-dependent transcription converting oxygen to hydrogen peroxide (might use alternative electron acceptors). Along with nuclear FHL2 it serves as a novel biomarker predictive for prostate cancer.

Alternate Names

AOF2, BHC110, LSD1, Lysine (K)specific Demethylase 1A

Gene Symbol

KDM1A

UniProt

Additional Lysine (K)-specific Demethylase 1A/KDM1A/LSD1 Products

Product Documents for LSD1 Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for LSD1 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

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Protocols

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Associated Pathways

Notch Signaling Pathways Notch Signaling Pathway Thumbnail