Spt6 Antibody

Western Blot: Spt6 Antibody [NB100-2582] - siRNA knockdown to confirm the specificity of Spt6 antibody (NB100-2582) in C2C12 myoblasts, WB: 1:2000 dilution. 20 ug whole cell lysate. WB image submitted by a verified customer review.

Western Blot: Spt6 Antibody [NB100-2582] - SPT6 Depletion Induces lncRNA Transcription(A) YWHAZ locus showing pre-mRNA (– strand) & PROMPT (+ strand). SPT6 depletion induced lncRNA as shown by ChRNA-seq & mNET-seq analyses. The profile of mNuc-seq/H3K36me3 is shown below.(B) Meta-analysis of strand-specific ChrRNA-seq signals from −3 kb of TSS to +3 kb of transcription end site (TES) for divergent (pre-mRNA-PROMPT) genes.(C) Boxplots of PROMPT mNET-seq/total CTD & T4P signals.(D) Enhancer located 90 kb downstream of NR4A1 gene with neighboring gene ATG101 showing SPT6 depletion-induced eRNA by ChRNA-seq & mNET-seq. mNuc-seq/H3K4me1 & H3K4me3 signals indicate active enhancer & promoter, respectively.(E) Meta-analysis of eRNA from ChrRNA-seq & mNET-seq (Total) −3 kb to +3 kb from TSS (enhancers with highest eRNA levels selected).(F) Boxplots of eRNA mNET-seq/total CTD & T4P signals at TSS (–/+2 kb).(G & H) Western blot (G) & quantitative RT-PCR (H) of chromatin-bound RNA of parental & SETD2 CRISPR KO U2OS cells with indicated siRNA transfection for 48 hr. Data are represented as mean ± SEM.See also Figure S3. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30449723), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Whole cell lysate (50 µg) from HEK293T, K-562,HeLa, TCMK-1, and NIH 3T3 cells prepared using NETNlysis buffer. Antibody: Affinity purified rabbit anti-SUPT6Hantibody used for WB at0.04 µg/ml. Detection: Chemiluminescence with anexposure time of 3 minutes.

Whole cell lysate (1.0 mgper IP reaction; 20% of IP loaded) from HEK293T cellsprepared using NETN lysis buffer. Antibodies: Affinitypurified rabbit anti-SUPT6H antibody) used for IP at 6 µg per reaction. SUPT6Hwas also immunoprecipitated by a previous lot of thisantibody. For blottingimmunoprecipitated SUPT6H, was used at 0.04µg/ml. Detection: Chemiluminescence with an exposuretime of 75 seconds.

Western Blot: Spt6 Antibody [NB100-2582] - SPT6 Depletion Causes H3K36me3 Redistribution(A) Western blots with indicated antibodies 60 hr post-siSPT6 transfection (versus siLuc nonspecific control). EXOSC3 & H3 profiles shown as loading controls.(B) Meta-analysis of reads density (FPKM) for ratio of mNuc-seq/H3K36me3 with H3 at TSS of divergent PROMPT-pre-mRNA & enhancer RNA (eRNA) following SPT6 depletion. All subsequent transcription images employing siSPT6 versus siLuc are shown in red & blue, respectively.(C) Scatterplots of H3K36me3/H3 on pre-mRNA (0 to +3 kb), PROMPT (−3 kb to 0), & eRNA (−3 kb to +3 kb) regions in siLuc versus siSPT6. The percentage of upregulated regions by SPT6 depletion are indicated.(D) Boxplots of mNuc-seq/H3K36me3 ratio across pre-mRNA gene bodies, PROMPTs (3 kb from TSS), & eRNA (2 kb from center) & across lincRNA gene bodies.(E) Model of redistributed H3K36me3 marks caused by SPT6 depletion.See also Figure S2. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30449723), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Section of human breast carcinoma.Antibody: Affinity purified rabbit anti- SUPT6H used at a dilution of 1:200 (1µg/ml).Detection: DAB

Section of mouse plasmacytoma. Antibody:Affinity purified rabbit anti- SUPT6H used at a dilution of 1:200 (1µg/ml). Detection: DAB

• Reactivity: Hu, Mu, Ch
• Applications: WB, IHC, IP, ChIP, Mycoplasma
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 0.2 mg/ml

Spt6 Antibody Summary

• Description: Novus Biologicals Rabbit Spt6 Antibody (NB100-2582) is a polyclonal antibody validated for use in IHC, WB, IP and ChIP. Anti-Spt6 Antibody: Cited in 19 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: The immunogen recognized by this antibody maps to a region between residue 425 and 475 of human Suppressor of Ty 6 Homolog using the numbering given in entry NP_003161.2 (GeneID 6830).
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: SUPT6H
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) Reported in scientific literature (PMID:23503590).
  • Immunohistochemistry 1:100 to 1:500
  • Immunohistochemistry-Paraffin 1:100 to 1:500
  • Immunoprecipitation 2 - 10 µg/mg lysate
  • Knockdown Validated
  • Western Blot 1:2000-1:10000
• Application Notes: Epitope retrieval with citrate buffer pH 6.0 is recommended for FFPE tissue sections.
• Control:

  HeLa Whole Cell Lysate

Reactivity Notes

Chicken reactivity reported in scientific literature (PMID: 23008333).

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: TBS, 0.1% BSA
• Preservative: 0.09% Sodium Azide
• Concentration: 0.2 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Spt6 Antibody

• emb-5
• hSPT6
• KIAA0162MGC87943
• SPT6HSPT6
• suppressor of Ty (S.cerevisiae) 6 homolog
• suppressor of Ty 6 homolog (S. cerevisiae)
• Tat-cotransactivator 2 protein
• Tat-CT2 protein
• transcription elongation factor SPT6

Background

Spt6 is a histone H3-H4 chaperone protein that mediates chromatin reassembly behind RNA polymerase II. Spt6 binds both histone H3 and H4 and deposits them on to DNA, which leads to transcriptional repression of target genes.

Limitations

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.

Publications for Spt6 Antibody (NB100-2582)(21)

Publications using NB100-2582

• Yu Z, Wang Q, Zhu G et al. Decoding the genomic landscape of chromatin-associated biomolecular condensates bioRxiv 2023-08-25 (WB)
• Gaballa A, Gebhardt-Wolf A, Krenz B et Al. PAF1c links S-phase progression to immune evasion and MYC function in pancreatic carcinoma Nat Commun 2024-02-16 [PMID: 38365788]
• Djakovic L, Hennig T, Reinisch K et al. The HSV-1 ICP22 protein selectively impairs histone repositioning upon Pol II transcription downstream of genes Nature Communications 2023-07-31 [PMID: 37524699]
• Lisa-Marie Appel, Vedran Franke, Johannes Benedum, Irina Grishkovskaya, Xué Strobl, Anton Polyansky, Gregor Ammann, Sebastian Platzer, Andrea Neudolt, Anna Wunder, Lena Walch, Stefanie Kaiser, Bojan Zagrovic, Kristina Djinovic-Carugo, Altuna Akalin, Dea Slade The SPOC domain is a phosphoserine binding module that bridges transcription machinery with co- and post-transcriptional regulators Nature Communications 2023-01-11 [PMID: 36631525]
• Liu X, Guo Z, Han J et al. The PAF1 complex promotes 3' processing of pervasive transcripts Cell reports 2022-03-15 [PMID: 35294889] (WB, Mouse)
• Narain A, Bhandare P, Adhikari B Et Al. Targeted protein degradation reveals a direct role of SPT6 in RNAPII elongation and termination Molecular cell 2021-06-30 [PMID: 34233157] (Chemotaxis)
• Obara E. A. A, Aguilar-Morante D, et al. SPT6-driven error-free DNA repair safeguards genomic stability of glioblastoma cancer stem-like cells. Nat Commun 2020-09-18 [PMID: 32948765] (Chemotaxis, Human)
• Lu H, Xie Y, Tran L et al. Chemotherapy-induced S100A10 recruits KDM6A to facilitate OCT4-mediated breast cancer stemness J Clin Invest. 2020-05-19 [PMID: 32427586] (IP, WB, KD, Human)
• Liming Hou, Yating Wang, Yu Liu, Nan Zhang, Ilya Shamovsky, Evgeny Nudler, Bin Tian, Brian David Dynlacht Paf1C regulates RNA polymerase II progression by modulating elongation rate Proceedings of the National Academy of Sciences of the United States of America 2019-07-16 [PMID: 31249142]
• Studniarek C, Tellier M, Martin PGP et al. The 7SK/P-TEFb snRNP controls ultraviolet radiation-induced transcriptional reprogramming Cell reports 2021-04-13 [PMID: 33852864]
• Chirackal Manavalan AP, Pilarova K, Kluge M et al. CDK12 controls G1/S progression by regulating RNAPII processivity at core DNA replication genes EMBO Rep. 2019-07-25 [PMID: 31347271] (IP, Human)
• Baluapuri A, Hofstetter J, Dudvarski Stankovic N et al. MYC Recruits SPT5 to RNA Polymerase II to Promote Processive Transcription Elongation Mol. Cell 2019-03-21 [PMID: 30928206] (WB, Mouse)
• Nojima T, Tellier M, Foxwell J et al. Deregulated Expression of Mammalian lncRNA through Loss of SPT6 Induces R-Loop Formation, Replication Stress, and Cellular Senescence. Toxicol. Sci. 2018-11-21 [PMID: 30449723] (Human)
• Nojima T, Rebelo k, Gomes T et al. RNA Polymerase II Phosphorylated on CTD Serine 5 Interacts with the Spliceosome during Co-transcriptional Splicing. Mol. Cell. 2018-10-18 [PMID: 30340024] (Human)
• Wang AH, Juan AH, Ko KD et al. The Elongation Factor Spt6 Maintains ESC Pluripotency by Controlling Super-Enhancers and Counteracting Polycomb Proteins. Mol. Cell. 2017-10-19 [PMID: 29033324] (WB)
• Wang AH, Zare H, Mousavi K et al. The histone chaperone Spt6 coordinates histone H3K27 demethylation and myogenesis. EMBO J 2013-03-15 [PMID: 23503590] (WB, Chemotaxis, Mouse)
• Willmann KL, Milosevic S, Pauklin S et al. A role for the RNA pol II-associated PAF complex in AID-induced immune diversification. J Exp Med. 2012-10-22 [PMID: 23008333] (IP, Chicken, Mouse)
• Chen S, Ma J, Wu F et al. The histone H3 Lys 27 demethylase JMJD3 regulates gene expression by impacting transcriptional elongation. Genes Dev 2012-06-01 [PMID: 22713873]
• Kronemann D, Hagemeier SR, Cygnar D et al. Binding of the human cytomegalovirus (HCMV) tegument protein UL69 to UAP56/URH49 is not required for efficient replication of HCMV. J Virol 2010-09-01 [PMID: 20610707]
• Hofstetter JEI MYC shapes the composition of RNA polymerase II through direct recruitment of transcription elongation factors Thesis 2022-01-01

Reviews for Spt6 Antibody (NB100-2582) (2)

Average Rating: 5

(Based on 2 reviews)

We have 2 reviews tested in 2 species: Human, Mouse.

reviewed by: Yongkang Yang WB Human 03/06/2021

Summary

• Application: Western Blot
• Sample Tested: breast cancer cell line, Sample Amount: 40
• Species: Human
• Lot: A2

reviewed by: Verified Customer WB Mouse 04/17/2018

Summary

• Application: Western Blot
• Sample Tested: 20 ug whole cell lysate,Mouse myoblasts (C2C12 cell lysate)
• Species: Mouse
• Lot: A1

Bioinformatics

• Gene Symbol: SUPT6H
• Entrez:
  • 6830(Human)
• Uniprot:
  • Q7KZ85(Human)