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Anti lsd1

Manufactured by Cell Signaling Technology
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Anti-LSD1 is a primary antibody that specifically recognizes and binds to the LSD1 (lysine-specific demethylase 1) protein. LSD1 is an enzyme that catalyzes the demethylation of histone H3 at lysine 4 and lysine 9, which are important epigenetic modifications involved in the regulation of gene expression.

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Lab products found in correlation

Anti h3k4me2, by Cell Signaling Technology (3 mentions) Anti h3k9me2, by Cell Signaling Technology (3 mentions) Anti gapdh, by Santa Cruz Biotechnology (2 mentions) Anti h3, by Cell Signaling Technology (2 mentions) Anti maml1, by Cell Signaling Technology (2 mentions) Anti cdk4, by Cell Signaling Technology (2 mentions) Anti gapdh, by Cell Signaling Technology (2 mentions) Anti β actin, by Cell Signaling Technology (2 mentions) Anti p21, by Cell Signaling Technology (2 mentions) Anti ezh2, by Cell Signaling Technology (2 mentions) Anti cd31, by Santa Cruz Biotechnology (2 mentions) Hx system, by Roche (2 mentions) Anti hes1, by Abcam (2 mentions) Anti runx2, by Cell Signaling Technology (2 mentions) Anti notch1, by Cell Signaling Technology (2 mentions) Anti gapdh, by ZSGB-BIO (2 mentions) Quantity one software, by Bio-Rad (1 mentions) Sds page, by Bio-Rad (1 mentions) Anti gapdh, by Proteintech (1 mentions) Hek293, by American Type Culture Collection (1 mentions)

Close spelling variants of this product

Rabbit anti-LSD1 (6 citations) Anti-LSD1 antibody (4 citations)

20 protocols using anti lsd1

1

Western Blot Analysis of HBP1 and LSD1

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Cell protein lysates were separated by SDS‐PAGE (Bio‐Rad, Hercules, CA, USA) and transferred to 0.45 μm PVDF transfer membranes (Thermo Fisher Scientific, Rockford, IL, USA). The membrane was blocked with 5% nonfat dry milk in 0.1% TBS‐Tween‐20 (TBST) for 2 hours, and then incubated with primary Abs for the night. The blots were washed with TBST three times for approximately 5 minutes a time, followed by incubation with HRP‐conjugated secondary Abs. Enhanced chemiluminescent HRP substrate was used for quantification by densitometry (Quantity One software; Bio‐Rad). Anti‐GAPDH (Proteintech Group, Chicago, IL, USA) was used as a control. Anti‐HBP1 was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti‐LSD1 was purchased from Cell Signaling Technology (Boston, MA, USA).
Yu S., Yang D., Ye Y., Liu P., Chen Z., Lei T., Pu J., Liu L, & Wang Z. (2019). Long noncoding RNA actin filament‐associated protein 1 antisense RNA 1 promotes malignant phenotype through binding with lysine‐specific demethylase 1 and repressing HMG box‐containing protein 1 in non‐small‐cell lung cancer. Cancer Science, 110(7), 2211-2225.
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2

Antibody Characterization and Cell Line Authentication

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Anti-HA, anti-tubulin and anti-GAPDH antibodies were purchased from Santa Cruz Biotechnology. Anti-ERK1/2, anti-pERK1/2 (Thr202/Tyr204), anti-Akt, anti-pAkt (Ser473) and anti-LSD1 antibodies were obtained from Cell Signaling. Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum, penicillin, and streptomycin were from Invitrogen. HEK293, MeWo and MCF7 cell lines were purchased directly from ATCC between 2008 and 2015. The ATCC cell lines were characterized by short tandem repeat (STR) DNA profiling. MCF10A cell was received as a gift from Dr. Mircea Ivan's lab in Indiana University School of Medicine, and was authenticated by morphology. All cell lines were passaged for fewer than 6 months after resuscitation.
Bai Y., Yu Z.H., Liu S., Zhang L., Zhang R.Y., Zeng L.F., Zhang S, & Zhang Z.Y. (2016). Novel anticancer agents based on targeting the trimer interface of the PRL phosphatase. Cancer research, 76(16), 4805-4815.
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3

Western Blot Antibody Validation

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Anti-LSD1 (Cell Signaling Technology, catalog 2139), anti-TP53 (Santa Cruz Biotechnology, catalog sc-126), anti-p21(CDKN1A) (Cell Signaling Technology, catalog 2947), anti-GAPDH (Santa Cruz Biotechnology, catalog sc-32233), anti-H3K4me2 (Cell Signaling Technology, catalog 9725), anti-H3K27Ac (Active Motif, catalog 39133), and anti–Histone H3 (MilliporeSigma, catalog 06-755) were used for protein detection by Western blotting.
Kumaraswamy A., Duan Z., Flores D., Zhang C., Sehrawat A., Hu Y.M., Swaim O.A., Rodansky E., Storck W.K., Kuleape J.A., Bedi K., Mannan R., Wang X.M., Udager A., Ravikumar V., Bankhead A I.I.I., Coleman I., Lee J.K., Morrissey C., Nelson P.S., Chinnaiyan A.M., Rao A., Xia Z., Yates J.A, & Alumkal J.J. (2023). LSD1 promotes prostate cancer reprogramming by repressing TP53 signaling independently of its demethylase function. JCI Insight, 8(15), e167440.
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4

Antibody Usage in Protein Detection

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The following commercially available antibodies were used: anti-Flag (Sigma), anti-HA (Roche), anti-LSD1 (Cell signaling), anti-RNA Polymerase II (Berkeley Antibody Company), anti-β-actin (Santa Cruz), anti-GFP (Santa Cruz), anti-Lamin A/C (Santa Cruz) and anti-GAPDH (Santa Cruz). Anti-RORα2 antibody (target epitope is GKPPYSQKEDKEVQT-C) was generated by Abmart (China).
Kim K., Lee J.M., Yu Y.S., Kim H., Nam H.J., Moon H.G., Noh D.Y., Kim K.I., Fang S, & Baek S.H. (2017). RORα2 requires LSD1 to enhance tumor progression in breast cancer. Scientific Reports, 7, 11994.
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5

Antibody-based Protein Detection Protocol

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The following antibodies were used for immunodetection: M2-anti-FLAG (HRP; Sigma A8592), anti-FLI-1 (Santa-Cruz sc-356X), anti-α-Tubulin (Calbiochem CP06), anti-HA (Abcam ab9110), anti-H3 total (Abcam ab1791), anti-H3K4 me1 (Abcam ab8895), anti-H3K4 me2 (Millipore, 07-030), anti-H3K4 me3 (Active Motif, 39159), anti-H3K9 me1 (Abcam ab9045), anti-H3K9 me2 (Abcam ab1220), anti-H3K9 me3 (Abcam ab8898), anti-HMOX1 (Sigma SAB1410641), anti-Paxillin (BD Transduction Labs 610619), anti-LSD1 (Cell Signaling Technology 2184) AlexaFluor secondary (Molecular Probes), AlexaFluor Phalloidin (Molecular Probes). HCI2509 is previously described (33 (link)).
Sankar S., Theisen E.R., Bearss J., Mulvihill T., Hoffman L.M., Sorna V., Beckerle M.C., Sharma S, & Lessnick S.L. (2014). Reversible LSD1 inhibition interferes with global EWS/ETS transcriptional activity and impedes Ewing sarcoma tumor growth. Clinical cancer research : an official journal of the American Association for Cancer Research, 20(17), 4584-4597.
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6

Immunodetection of Epigenetic Markers

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Immunodetection was performed with the following antibodies: anti-α-Tubulin (Calbiochem CP06), anti-LSD1 (Cell Signaling C69G12), anti-H3 (Cell Signaling Technology D2B12), anti-H3K4me3 (Cell Signaling Technology C42D8), anti-H3K9me2 (Cell Signaling Technology 9753), anti-H3K27me3 (Cell Signaling Technology C36B11). Propidium iodide (Sigma P4864), medroxyprogesterone 17-acetate (MPA; Sigma M1629). HCI2509 is previously described [35 (link)].
Theisen E.R., Gajiwala S., Bearss J., Sorna V., Sharma S, & Janat-Amsbury M. (2014). Reversible inhibition of lysine specific demethylase 1 is a novel anti-tumor strategy for poorly differentiated endometrial carcinoma. BMC Cancer, 14, 752.
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7

Investigating Notch Signaling Cascades

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Retroviral Infections, qRT-PCR Analysis, nuclear extraction and Western Blotting were performed as described previously (26 ,27 (link)). Cell samples were collected 72h post-infection. The following antibodies were used in Western blotting: anti-Notch1 (1:1000, abcam), anti-CSL (1:1000; polyclonal), anti-MAML1 (1:1000, cell signaling), anti-LSD1 (1:1000, cell signaling) and anti-EZH2 (1:1000, cell signaling).
Han X., Ranganathan P., Tzimas C., Weaver K.L., Jin K., Astudillo L., Zhou W., Zhu X., Li B., Robbins D.J, & Capobianco A.J. (2017). Notch Represses Transcription by PRC2 Recruitment to the Ternary Complex. Molecular cancer research : MCR, 15(9), 1173-1183.
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8

Western Blotting Analysis of Tumor Proteins

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Western blotting was performed according to the standard protocol with the protein lysates harvested form fresh tumor samples and cultured cells. Two µg of total protein was applied to one end of a 12% SDS polyacrylamide gel. After electrophoresis proteins on the gel were transferred onto a nitrocellulose membrane (Millipore, Temecula, CA, USA). After blocking with non-fat milk for almost 1 h at room temperature, the membranes were incubated overnight at 4 °C with primary antibodies: anti-LSD1 (1:1000), anti-H3K4me2 (1:1000), anti-H3K9me2 (1:1000), anti-P53(1:500), anti-P21 (1:500), anti-CDK4 (1:1000), anti-CDK6 (1:1000), anti-GAPDH (1:2000) and anti-β-actin (1:1000), which were all purchased from Cell Signaling Technology (Boston, Massachusetts, USA). Membranes were washed three times and incubated with secondary antibodies (1:500; Abcam, Cambridge, MA, USA) at room temperature for 1 h. Immunoreactive bands were detected by using Amersham Hyper lm ECL (GE Healthcare Life Sciences). GAPDH and β-actin were used as loading controls.
Zhu L., Wang J., Kong W., Huang J., Dong B., Huang Y., Xue W, & Zhang J. (2018). LSD1 inhibition suppresses the growth of clear cell renal cell carcinoma via upregulating P21 signaling. Acta Pharmaceutica Sinica. B, 9(2), 324-334.
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9

Western Blot Analysis of Epigenetic Regulators

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Protein lysates were prepared as previously described [38 (link)]. The protein samples were resolved by SDS-PAGE and transferred onto PVDF membranes (Roche, Basel, Switzerland). The membranes were then incubated with the following antibodies: anti-LSD1 (Cell Signaling Technology), anti-EZH2 (Cell Signaling Technology), anti-p16 (Santa Cruz), anti-p21 (Santa Cruz), anti-Cyclin D1 (Santa Cruz), anti-Cyclin E (Santa Cruz), anti-H3K27Me3 (Millipore), and anti-H3K4Me3 (Millipore). Antibody-labeled protein bands on the PVDF membranes were detected using a G:BOX F3 (Syngene, Cambridge, UK).
Zhang K., Sun X., Zhou X., Han L., Chen L., Shi Z., Zhang A., Ye M., Wang Q., Liu C., Wei J., Ren Y., Yang J., Zhang J., Pu P., Li M, & Kang C. (2014). Long non-coding RNA HOTAIR promotes glioblastoma cell cycle progression in an EZH2 dependent manner. Oncotarget, 6(1), 537-546.
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10

Western Blot Analysis of Liver Proteins

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Liver samples were homogenized in RIPA buffer (20 mM Tris-Cl, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% IGEPAL, and 1% deoxycholate) supplemented with Halt protease/phosphatase inhibitor (Thermo Fisher Scientific), centrifuged for 10 minutes at 10 000× g and protein concentration in the supernatants measured by the BCA method (Thermo Fisher Scientific). Fifty μg protein was separated on CriterionTGX Any kD precast gels (Bio-Rad Laboratories, Hercules, CA) and electroblotted onto Immun-Blot PVDF membranes (Bio-Rad Laboratories). Blots were incubated overnight at 4°C with the indicated primary antibodies (see below), and then, for 1 hour with HRP-conjugated secondary antibodies (Thermo Fisher Scientific). Signals were developed with Clarity Max Western ECL substrate and collected using the ChemiDoc imaging system and quantitated with Image Lab software, version 6.0.1 (all from Bio-Rad Laboratories). Primary antibodies used: anti-FRA-1 (catalog no. PA5–76185; Thermo Fisher Scientific), anti-IL-1β (catalog no. ab9722; Abcam, Cambridge, MA), anti-phospho-JNK1 (catalog no. 9255S; Cell Signaling Technology, Danvers, MA), anti-JNK1 (catalog no. 370; BioVision, Milpitas, CA), anti-LSD1 (catalog no. 2139; Cell Signaling Technology), and anti-β-actin (catalog no. 4970; Cell Signaling Technology).
Warner D.R., Warner J.B., Hardesty J.E., Song Y.L., Chen C.Y., Chen Z., Kang J.X., McClain C.J, & Kirpich I.A. (2021). Beneficial effects of an endogenous enrichment in n3-PUFAs on Wnt signaling are associated with attenuation of alcohol-mediated liver disease in mice. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 35(2), e21377.
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