Anti nelf e

Manufactured by Santa Cruz Biotechnology

Sourced in Denmark

Anti-NELF-E is a laboratory reagent used in research applications. It is an antibody that specifically binds to the NELF-E protein, which is a component of the negative elongation factor (NELF) complex involved in the regulation of transcription. The core function of Anti-NELF-E is to enable the detection and study of NELF-E in biological samples.

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Lab products found in correlation

Dynabeads protein g, by Thermo Fisher Scientific (1 mentions) Anti mouse igg peroxidase conjugate, by Merck Group (1 mentions) Dynabeads protein a, by Thermo Fisher Scientific (1 mentions) Anti rabbit igg peroxidase conjugate, by Merck Group (1 mentions) Anti gapdh, by Fortis Life Sciences (1 mentions) Anti flag, by Merck Group (1 mentions) Alexa fluor 488 anti mouse, by Jackson ImmunoResearch (1 mentions) Alexafluor 594 anti rabbit, by Jackson ImmunoResearch (1 mentions) Anti p p38, by Cell Signaling Technology (1 mentions) D8l4y, by Abcam (1 mentions) Anti top2b, by Proteintech (1 mentions) Anti top2a, by Abcam (1 mentions) Sc 365916, by Abcam (1 mentions) Anti top2a, by Santa Cruz Biotechnology (1 mentions) Anti γh2ax, by Merck Group (1 mentions) Anti tubulin, by Merck Group (1 mentions) Anti hla a antibody, by Abcam (1 mentions) Anti myc, by Santa Cruz Biotechnology (1 mentions) Envision kit, by Agilent Technologies (1 mentions) Ab70550, by Abcam (1 mentions)

5 protocols using anti nelf e

Chromatin Immunoprecipitation (ChIP) Assay

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ChIP experiments were carried out as previously described (16 (link)). Cells were treated with 1% formaldehyde for 15 min at 37ºC for crosslinking. Immunoprecipitations (IPs) were performed using the following reagents: Total RNAPII: Dynabeads Protein A (Invitrogen) and rabbit polyclonal anti-RPB1 N-20 (Santa Cruz); RPB1 CTD-Ser5P: Dynabeads Protein G (Invitrogen) and anti-RNA polymerase II (phospho-CTD-Ser5) antibody clone 3E8 (Millipore); NELF-E: Dynabeads Protein A (Invitrogen) and rabbit polyclonal anti-NELF-E (Santa Cruz). Primers used are listed in Supplementary Table S1.
Antibodies used for Western blotting are rabbit polyclonal anti-NELF-E (sc-32912, Santa Cruz Biotech.), rabbit polyclonal anti-SNF2H (A301–017A, Bethyl), mouse monoclonal anti-α-Tubulin and, as secondary antibodies, anti-Rabbit IgG peroxidase conjugate (Sigma) and anti-Mouse IgG peroxidase conjugate (Sigma).

Jimeno-González S., Ceballos-Chávez M, & Reyes J.C. (2015). A positioned +1 nucleosome enhances promoter-proximal pausing. Nucleic Acids Research, 43(6), 3068-3078.

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Granulocyte Protease Inhibition for Immunoblotting

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Differentiating granulocytes were first incubated in 5.4mM diisopropyl fluorophosphate (DFP) for 15 minutes on ice to prevent protein degradation caused by neutrophil-derived protease^{16 (link)}. Washed cells were then lysed directly in 1x SDS loading buffer followed by immediate boiling in the presence of 100mM DTT for 10 minutes before loading on SDS-PAGE for Western blotting. Antibodies used are: Anti-GAPDH (Bethyl, A300-641A), Anti-NELFA (Bethyl, A301-910A), Anti-NELFB (Bethyl, A301-912A), Anti-NELFD (Santa Cruz, sc393972), Anti-NELFE (Santa Cruz, sc377052), anti-Flag (Sigma, F1804).

Liu X., Gogate A.A., Tastemel M., Malladi V.S., Yao H., Nguyen K., Huang L.J, & Bai X. (2017). Dynamic change of transcription pausing through modulating NELF protein stability regulates granulocytic differentiation. Blood Advances, 1(18), 1358-1367.

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Topoisomerase-Dependent Genomic Interactions

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For ChIP-seq we have used anti-Rpb1-NTD (Cell Signaling, D8L4Y), anti-TOP2A (Abcam, ab52934). For ICE, anti-TOP2B (Proteintech, 20549-1-AP), anti-TOP2A (Santa Cruz, SC-365916), anti-TOP1 (Abcam, ab3825). For ICE-IP, anti-TOP2A (abcam, ab52934), anti-TOP1 antibody (abcam, ab109374). For western blot analysis, anti-TOP2B (Proteintech, 20549-1-AP), anti-TOP2A (Santa Cruz, SC-365916), anti-p-p38 (Cell Signaling, 9211) and anti-tubulin (Sigma, T9026), and as secondary antibodies IRDye 680-labeled anti-mouse (LI-COR Biosciences, 926-68070) and IRDye 800-labeled anti-rabbit (LI-COR BIOSCIENCES, 926-32211).
For immunofluorescence, anti-γH2AX (Millipore, 05-636) and anti-H3S10p (Millipore, 06-570). For PLA, anti-NELF-E (Santa Cruz, sc32912), anti-TOP2A (Santa Cruz, SC-365916), and as secondary antibodies Alexa Fluor 488 anti-mouse (Jackson, 715-545-150) and Alexa Fluor 594 anti-rabbit (Jackson, 111-585-003).

Herrero-Ruiz A., Martínez-García P.M., Terrón-Bautista J., Millán-Zambrano G., Lieberman J.A., Jimeno-González S, & Cortés-Ledesma F. (2021). Topoisomerase IIα represses transcription by enforcing promoter-proximal pausing. Cell Reports, 35(2), 108977.

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Immunohistochemical Analysis of HLA-A, MYC, and NELFE

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The paraffin blocks were sectioned at 5 μm and stained with haematoxylin and eosin. Anti-HLA-A antibody (Abcam), anti-MYC (Santa Cruz), anti-NELFE (Santa Cruz), and and Envision™+ Kit (Dako, Glostrup, Denmark) were used for detection.

Dang H., Takai A., Forgues M., Pomyen Y., Mou H., Xue W., Ray D., Ha K.C., Morris Q.D., Hughes T.R, & Wang X.W. (2017). Oncogenic activation of the RNA binding protein NELFE and MYC signaling in hepatocellular carcinoma. Cancer cell, 32(1), 101-114.e8.

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SUMO2 Antibody Production and Validation

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SUMO hybridoma SUMO2 8A2, were procured from Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology (http://dshb.biology.uiowa.edu/) and supernatant was produced in Pichler lab. The SUMO2 antisera were used at 1:100. All other primary antibodies were purchased and validated by the manufacturers (data available on manufacturers’ websites). They were used at the indicated dilutions; anti-NELFA (goat polyclonal, sc-23599 (A-20), Santa Cruz Biotechnology) at 1:1,000, anti-NELFC/D (mouse monoclonal, sc-393972 (C-10), Santa Cruz Biotechnology) 1:1,000, anti-NELFE (rabbit polyclonal, sc-32912 (H-140), Santa Cruz Biotechnology) at 1:1,000, anti-SNRNP70 (rabbit polyclonal, ab83306, Abcam) at 1:1,000, anti-ZNF451 (rabbit polyclonal, A305-177A, Bethyl Laboratories inc), anti-RPB3 (rabbit polyclonal, ABE999, Millipore) at 1:1,000, and anti-Histone H3 (rabbit polyclonal, ab70550, Abcam) at 1:5,000. Secondary antibodies against goat, rabbit or mouse proteins were horseradish peroxidase conjugated and were typically used at 1:10,000 dilution. Anti-rabbit (NA934V) and anti-mouse (NA931V) antibodies were purchased from GE healthcare. Anti-goat (sc-2354) antibody was purchased from Santa Cruz Biotechnology.

Rawat P., Boehning M., Hummel B., Aprile-Garcia F., Pandit A.S., Eisenhardt N., Khavaran A., Niskanen E., Vos S.M., Palvimo J.J., Pichler A., Cramer P, & Sawarkar R. (2021). Stress-induced nuclear condensation of NELF drives transcriptional downregulation. Molecular Cell, 81(5), 1013-1026.e11.

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