Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Spt6 antibody (ab32820)

Key features and details

  • Rabbit polyclonal to Spt6
  • Suitable for: IP, WB
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-Spt6 antibody
    See all Spt6 primary antibodies

  • Description

    Rabbit polyclonal to Spt6

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide corresponding to Human Spt6 aa 1150-1250 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab32818, ab32819)

  • Positive control

    • This antibody gave a positive signal for the following cell lysates: HeLa Nuclear; HeLa; Jurkat.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration
  • Concentration information loading...

  • Purity

    Immunogen affinity purified

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32820 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
IP
Human
WB
Human
All applications
Mouse
Zebrafish
Application Abreviews Notes
IP
Use at an assay dependent concentration.
WB (1)
1/500. Detects a band of approximately 230 kDa (predicted molecular weight: 199 kDa).
Notes
IP
Use at an assay dependent concentration.
WB
1/500. Detects a band of approximately 230 kDa (predicted molecular weight: 199 kDa).

Target

Images

  • Western blot - Anti-Spt6 antibody (ab32820)
    Western blot - Anti-Spt6 antibody (ab32820)
    Lane 1 : Marker
    Lanes 2-4 : Anti-Spt6 antibody (ab32820) at 1/500 dilution
    Lane 5 : High Mark marker

    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
    Lane 3 : Jurkat whole cell lysate (ab7899) at 20 µg
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 20 µg

    Secondary
    Lanes 2-4 : IR Dye 680 Conjugated Goat anti Rabbit IgG (H+L) at 1/21250 dilution

    Performed under reducing conditions.

    Predicted band size: 199 kDa
    Observed band size: 230 kDa
    why is the actual band size different from the predicted?



    Ab32820 detects a band at approximately 230kDa.  Other antibodies to this target detect a band in this region and it is understood that this band corresponds to the native Spt6 protein.
  • Immunoprecipitation - Anti-Spt6 antibody (ab32820)
    Immunoprecipitation - Anti-Spt6 antibody (ab32820)
    Spt6 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Spt6 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32820.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 230kDa: Spt6

References (9)

Publishing research using ab32820? Please let us know so that we can cite the reference in this datasheet.

ab32820 has been referenced in 9 publications.

  • Soboleva TA  et al. A new link between transcriptional initiation and pre-mRNA splicing: The RNA binding histone variant H2A.B. PLoS Genet 13:e1006633 (2017). ChIP ; Mouse . PubMed: 28234895
  • Dembowski JA  et al. Replication-Coupled Recruitment of Viral and Cellular Factors to Herpes Simplex Virus Type 1 Replication Forks for the Maintenance and Expression of Viral Genomes. PLoS Pathog 13:e1006166 (2017). PubMed: 28095497
  • Gérard A  et al. The integrase cofactor LEDGF/p75 associates with Iws1 and Spt6 for postintegration silencing of HIV-1 gene expression in latently infected cells. Cell Host Microbe 17:107-17 (2015). PubMed: 25590759
  • Dembowski JA & DeLuca NA Selective recruitment of nuclear factors to productively replicating herpes simplex virus genomes. PLoS Pathog 11:e1004939 (2015). ICC/IF . PubMed: 26018390
  • Dietrich JE  et al. Venus trap in the mouse embryo reveals distinct molecular dynamics underlying specification of first embryonic lineages. EMBO Rep 16:1005-21 (2015). PubMed: 26142281
  • Nakamura M  et al. Spt6 levels are modulated by PAAF1 and proteasome to regulate the HIV-1 LTR. Retrovirology 9:13 (2012). ICC/IF . PubMed: 22316138
  • Gallastegui E  et al. Chromatin Reassembly Factors Are Involved in Transcriptional Interference Promoting HIV Latency. J Virol 85:3187-202 (2011). WB . PubMed: 21270164
  • Vanti M  et al. Yeast genetic analysis reveals the involvement of chromatin reassembly factors in repressing HIV-1 basal transcription. PLoS Genet 5:e1000339 (2009). WB ; Human . PubMed: 19148280
  • Shen X  et al. Identification of novel SHPS-1-associated proteins and their roles in regulation of insulin-like growth factor-dependent responses in vascular smooth muscle cells. Mol Cell Proteomics : (2009). WB . PubMed: 19299420

Images

  • Western blot - Anti-Spt6 antibody (ab32820)
    Western blot - Anti-Spt6 antibody (ab32820)
    Lane 1 : Marker
    Lanes 2-4 : Anti-Spt6 antibody (ab32820) at 1/500 dilution
    Lane 5 : High Mark marker

    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
    Lane 3 : Jurkat whole cell lysate (ab7899) at 20 µg
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 20 µg

    Secondary
    Lanes 2-4 : IR Dye 680 Conjugated Goat anti Rabbit IgG (H+L) at 1/21250 dilution

    Performed under reducing conditions.

    Predicted band size: 199 kDa
    Observed band size: 230 kDa
    why is the actual band size different from the predicted?



    Ab32820 detects a band at approximately 230kDa.  Other antibodies to this target detect a band in this region and it is understood that this band corresponds to the native Spt6 protein.
  • Immunoprecipitation - Anti-Spt6 antibody (ab32820)
    Immunoprecipitation - Anti-Spt6 antibody (ab32820)
    Spt6 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Spt6 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32820.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 230kDa: Spt6

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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