Product nameAnti-Spt6 antibody
DescriptionRabbit polyclonal to Spt6
Tested applicationsSuitable for: IHC-P, IP, WBmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Zebrafish
- This antibody gave a positive signal for the following cell lysates: HeLa Nuclear; HeLa; Jurkat.
Storage instructionsShipped 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 bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS. pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
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.
|IHC-P||1/30. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IP||Use at an assay dependent concentration.|
|WB||1/500. Detects a band of approximately 230 kDa (predicted molecular weight: 199 kDa).|
FunctionActs to stimulate transcriptional elongation by RNA polymerase II.
Tissue specificityUbiquitously expressed.
Sequence similaritiesBelongs to the SPT6 family.
Contains 1 S1 motif domain.
Contains 1 SH2 domain.
- Information by UniProt
- emb 5 antibody
- hSPT6 antibody
- KIAA0162 antibody
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
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.
Image courtesy of Human Protein Atlas
ab32820 staining Spt6 in human gall bladder, showing a distinct and strong staining pattern in glandular cells. Paraffin embedded human skin tissue was incubated with ab32820 (1/30 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6..
ab32820 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
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
This product has been referenced in:
- 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 . Read more (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). Read more (PubMed: 28095497) »