Key features and details
- Rabbit polyclonal to ISWI (acetyl K753)
- Suitable for: WB
- Reacts with: Drosophila melanogaster, Recombinant fragment
- Isotype: IgG
Product nameAnti-ISWI (acetyl K753) antibody
DescriptionRabbit polyclonal to ISWI (acetyl K753)
SpecificityThe specificity of the antibody can be summarized as follows: 1. On Western blots probing recombinant ISWI which had been (or not) acetylated at K753 by GCN5, the antibody recognizes the acetylated ISWI with very high selectivity. No reaction with unacetylated ISWI has been observed. 2. Unexpectedly, there is a significant cross-reactivity with GCN5 under acetylation conditions (we assume autoacetylated GCN5). As shown in the figure, this cross-reactivity is low, since conditions can be found easily where the GCN5 band on Western blots is not observed. 3. In whole cell extracts the antibody cross-reacts with a number of small molecular weight bands. A faint signal at the position of ISWI is detected as well, which we think corresponds to ISWI since it is not present if ISWI is ablated using RNAi. The weakness of the signal may simply indicate that there is very little epitope present in these cells. In summary, we are confident that the antibody can be used on purified ISWI (e.g. ISWI immunoprecipitated from a cruder source).
Tested applicationsSuitable for: WBmore details
Species reactivityReacts with: Drosophila melanogaster, Recombinant fragment
Predicted to work with: Sea urchin
Synthetic peptide corresponding to Drosophila melanogaster ISWI aa 700-800 conjugated to keyhole limpet haemocyanin.
(Peptide available as
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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
PurityImmunogen affinity purified
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab10748 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/2500. Detects a band of approximately 110 kDa (predicted molecular weight: 119 kDa).Can be blocked with D. melanogaster ISWI (acetyl K753) peptide (ab16064).|
RelevanceISWI is a component of the nucleosome remodeling factor complex (NURF), a protein complex that facilitates the perturbation of chromatin structure in vitro in an ATP-dependent manner. The hydrolysis of ATP during the remodeling of chromatin is likely to be mediated by ISWI, releasing inorganic phosphate. It is also a component of the ATP-utilizing chromatin assembly and remodeling factor (ACF) and of the chromatin accessibility complex (CHRAC). This subunit may serve as the energy-transducing component of chromatin-remodeling machines.
- CHRAC 140 kDa subunit antibody
- dISWI antibody
- Imitation SWI antibody
Lanes 1-2 : Anti-ISWI (acetyl K753) antibody (ab10748) at 1/2500 dilution
Lanes 3-4 : Anti-ISWI (acetyl K753) antibody (ab10748) at 1/500 dilution
Lanes 2-3 : Recombinant ISWI (200 ng) was acetylated using 125ng of hGCN5 and a final concentration of 6 µM of 3H-AcCoA.
Predicted band size: 119 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?
Additional bands at: 65 kDa. We are unsure as to the identity of these extra bands.
All lanes : Anti-ISWI (acetyl K753) antibody (ab10748) at 1/500 dilution
Lane 1 : Drosophila melanogaster SL-2 cells treated for 8 days with ISWI dsRNAs
Lane 2 : Drosophila melanogaster SL-2 cells treated for 8 days with control dsRNAs
Lysates/proteins at 30 µg per lane.
Predicted band size: 119 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?
Additional bands at: 55 kDa, 60 kDa, 65 kDa. We are unsure as to the identity of these extra bands.
ab10748 has been referenced in 4 publications.
- L Black K et al. Chromatin proteins and RNA are associated with DNA during all phases of mitosis. Cell Discov 2:16038 (2016). PubMed: 27807477
- Yajima M et al. ISWI contributes to ArsI insulator function in development of the sea urchin. Development 139:3613-22 (2012). ICC/IF ; Sea urchin . PubMed: 22949616
- Li M et al. Modulation of chromatin boundary activities by nucleosome-remodeling activities in Drosophila melanogaster. Mol Cell Biol 30:1067-76 (2010). ChIP ; Drosophila melanogaster . PubMed: 19995906
- Ferreira R et al. Site-specific acetylation of ISWI by GCN5. BMC Mol Biol 8:73 (2007). PubMed: 17760996