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Purified anti-Spt16 (FACT140 complex) Antibody

Pricing & Availability
Clone
8D2 (See other available formats)
Regulatory Status
RUO
Other Names
Facilitates Chromatin Transcription (FACT140), Spt 16 protein, Cdc68
Isotype
Mouse IgG2a, κ
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Product Citations
publications
1_8D2_Spt16(FACT140)_Purfied_antibody_WB_060414.jpg
Total lysates (15 µg protein) from A549 (lane 1), Jurkat (lane 2), and Raw 264.7 cells (lane 3) were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with 1:500 diluted (1 µg/mL) Purified anti-Spt16 Antibody, clone 8D2 (upper). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for the anti-Spt16 Antibody or 1:5000 diluted Direct-Blot HRP anti-β-Actin Antibody, clone 2F1-1(lower). Lane M: Molecular weight ladder.
  • 1_8D2_Spt16(FACT140)_Purfied_antibody_WB_060414.jpg
    Total lysates (15 µg protein) from A549 (lane 1), Jurkat (lane 2), and Raw 264.7 cells (lane 3) were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with 1:500 diluted (1 µg/mL) Purified anti-Spt16 Antibody, clone 8D2 (upper). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for the anti-Spt16 Antibody or 1:5000 diluted Direct-Blot HRP anti-β-Actin Antibody, clone 2F1-1(lower). Lane M: Molecular weight ladder.
  • 2_Z_8D2_Spt16_Purified_Antibody_IF_011515
    Hela cells were fixed with 2% paraformaldehyde (PFA) for 10 minutes, permeabilized with 0.5% Triton X-100 for 5 minutes, and blocked with 5% FBS for 30 minutes. Then the cells were intracellularly stained with 5 µg/mL anti-Spt16 Antibody (8D2) and followed by DyLight™ 594 goat anti-mouse IgG (red) for one hour at room temperature. Actin filaments were labeled with Alexa Fluor® 488 Phalloidin (green). Nuclei were counterstained with DAPI (blue). The image was captured with a 60X objective.
  • 3_8D2_GoChIP_Spt16_FACT140_Antibody_1_052118.png
    Chromatin Immunoprecipitations (ChIP) were performed with cross-linked chromatin samples from 4 X 106 HT29 cells with either A) 1:50 dilution of Go-ChIP-Grade™ Purified anti-Spt16 (FACT140 complex) (clone 8D2, Cat. No. 607007) or B) equal amount of Go-ChIP-Grade™ Purified Mouse IgG2a, κ isotype control antibody (clone MG2a-53, Cat. No. 401505) by using Go-ChIP-Grade™ Protein G Enzymatic Kit (Cat. No. 699904). The enriched DNA was purified and quantified by real-time qPCR using primers targeting human EGR1 gene region or α-Satellite repeats. The amount of immunoprecipitated DNA in each sample is represented as signal relative to total amount of input chromatin.
  • 4_8D2_GoChIP_Spt16_FACT140_Antibody_2_052118.PNG
    Chromatin Immunoprecipitations (ChIP) were performed with cross-linked chromatin samples from 4 X 106 HT29 cells with either A) 1:50 dilution of Go-ChIP-Grade™ Purified anti-Spt16 (FACT140 complex) (clone 8D2, Cat. No. 607007) or B) equal amount of Go-ChIP-Grade™ Purified Mouse IgG2a, κ isotype control antibody (clone MG2a-53, Cat. No. 401505) by using Go-ChIP-Grade™ Protein G Enzymatic Kit (Cat. No. 699904). The enriched DNA was purified and quantified by real-time qPCR using primers targeting human C-FOS gene region or α-Satellite repeats. The amount of immunoprecipitated DNA in each sample is represented as signal relative to total amount of input chromatin.
Cat # Size Price Quantity Check Availability Save
607001 25 µg £70
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607002 100 µg £150
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Description

Spt16 is a 120-140 kD nuclear protein and a component of the FACT complex (facilitates chromatin transcription). The FACT140 complex is composed of the Spt16 and the SSRP1 proteins. This complex interacts with nucleosomes and histone H2A/H2B dimers to promote nucleosome disassembly and allow transcription elongation. The 8D2 monoclonal antibody recognizes human Spt16 and has been shown to be useful for Western blotting.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Recombinant (full length)
Formulation
This antibody is provided in phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
Upon receipt, store between 2°C and 8°C.
Application

WB - Quality tested
ICC, ChIP - Verified
IP - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. Western blotting, suggested working dilution(s): Use 5 µg per 5 ml antibody dilution buffer for each mini-gel. For immunocytochemistry, a concentration range of 1.0 - 5.0 µg per ml is recommended. For ChIP applications, the suggested dilution is 1:50 - 1:100 by volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

25 µg, 100 µg of Go-ChIP-Grade™ Purified Antibody can be used for 2-4, 11-16 immunoprecipitations, respectively, at the recommended dilution. ICC positive in HeLa Cells. 

Application References

(PubMed link indicates BioLegend citation)
  1. Tan B C-M, et al. 2006. EMBO doi:10.1038/sj.emboj.7601271
  2. Adelmant G, et al. 2012. Mol Cell Proteomics. 11:411. PubMed
  3. Lolis AA, et al. 2013. J Biol Chem. 288:7676. PubMed
  4. Shakya A, et al. 2015. Mol Cell Biol. 35:1014. PubMed
Product Citations
  1. Goswami I, et al. 2022. EMBO Rep. 23:e53684. PubMed
  2. Burmistrova D, et al. 2016. PLoS One. 11: 0150958. PubMed
  3. Fleyshman D, et al. 2017. Oncotarget. 8:20525. PubMed
  4. Adelmant G, et al. 2012. Mol Cell Proteomics. 11:411. PubMed
  5. Isa NF, et al. 2021. Vaccines (Basel). 9:. PubMed
  6. Safina A, et al. 2017. Nucleic Acids Res. 10.1093/nar/gkw1366. PubMed
  7. Shakya A, et al. 2015. Mol Cell Biol. 35:1014. PubMed
  8. Hu X, et al. 2019. Front Genet. 10:684. PubMed
  9. Kaja A, et al. 2021. Mol Cell Biol. 41:. PubMed
  10. Lolis A, et al. 2013. J Biol Chem. 288:7676. PubMed
  11. Lin D, et al. 2018. EMBO J. 37. PubMed
  12. Sandlesh P, et al. 2018. PLoS One. 15:81. PubMed
  13. Prendergast L, et al. 2020. Oncogene. 39:5124. PubMed
RRID
AB_315688 (BioLegend Cat. No. 607001)
AB_315688 (BioLegend Cat. No. 607002)

Antigen Details

Structure
120-140 kD
Distribution

Nuclear

Function
Interacts with nucleosomes and histone H2A/H2B dimers to promote nucleosome disassembly and allow transcription elongation
Interaction
Component of FACT ATP-dependent chromatin remodeling complex, SSRP1 protein in FACT complex, H2A/H2B dimers
Biology Area
Cell Biology, Chromatin Remodeling/Epigenetics, Transcription Factors
Molecular Family
Nuclear Markers
Antigen References

1. LeRoy G, et al. 1998. Science 282:1836.
2. Orphanides G, et al. 1998. Cell 92:105.
3. Orphanides G, et al. 1999. Nature 400:284.

Gene ID
11198 View all products for this Gene ID
UniProt
View information about Spt16 FACT140 complex on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 6    Revision Date: 08/24/2021

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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