Product nameAnti-ISWI (acetyl K753) antibody
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).
Species reactivityReacts with: Drosophila melanogaster, Sea urchin
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
Immunizing Peptide (Blocking)
|ELISA||Use at an assay dependent concentration. This antibody gave a positive result in ELISA against the immunizing peptide (ab16064). It gave a negative result in ELISA against the non-modified equivalent peptide (ab96245). This indicates that it is specific for the modified peptide.|
|ICC/IF||Use at an assay dependent concentration. PubMed: 22949616|
|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.
This product has been referenced in:
- L Black K et al. Chromatin proteins and RNA are associated with DNA during all phases of mitosis. Cell Discov 2:16038 (2016). Read more (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 . Read more (PubMed: 22949616) »