This fast track antibody is not yet fully characterized. It is subject to these terms and conditions

Overview

  • Product nameAnti-ISWI (acetyl K753) antibody
  • Description
    Rabbit polyclonal to ISWI (acetyl K753)

    This product is a fast track antibody. It has been affinity purified and shows high titre values against the immunizing peptide by ELISA. Read the terms of use »

  • 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 reactivity
    Reacts with: Drosophila melanogaster, Sea Urchin
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 700 - 800 of Fruit fly (Drosophila melanogaster) ISWI, acetylated at K753.

    (Peptide available as ab16064.)

Properties

  • FormLiquid
  • 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
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Associated products

Applications

Fast track antibodies constitute a diverse group of products that have been released to accelerate your research, but are not yet fully characterized. They have all been affinity purified and show high titre values against the immunizing peptide (by ELISA). Fast track terms of use

Application Abreviews Notes
ELISA Use at an assay dependent dilution. 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

Target

  • 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.
  • Cellular localizationNuclear
  • Database links
    • Alternative names
      • CHRAC 140 kDa subunit antibody
      • dISWI antibody
      • Imitation SWI antibody
      • NURF 140 antibody
      • NURF antibody
      • Protein imitation swi antibody
      see all

    Anti-ISWI (acetyl K753) antibody images

    This Fast-Track antibody is not yet fully characterised. These images represent inconclusive preliminary data.

    • 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

      Lane 1 : As above
      Lane 2 : Recombinant ISWI (200 ng) was acetylated using 125ng of hGCN5 and a final concentration of 6 µM of 3H-AcCoA.
      Lane 3 : Recombinant ISWI (200 ng) was acetylated using 125ng of hGCN5 and a final concentration of 6 µM of 3H-AcCoA.
      Lane 4 : As above


      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.

      This image is courtesy of Peter Becker

      Recombinant ISWI (200 ng) was acetylated using 125 ng (lanes 2 and 3) of hGCN5 and a final concentration of 6 µM of 3H-AcCoA.

      Acetylated proteins were detected by western blotting using the acetylated ISWI (K753) polyclonal antibody (ab10748), diluted 1/2500 (lanes1 and 2) or 1/500 (lanes 3 and 4) in 3% milk 0.1% T-PBS. In lanes 3 and 4, 1/5th of the reaction was loaded.

       

      In lanes 3 and 4, 1/5th of the reaction was loaded.
    • 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.


      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 image is courtesy of Peter Becker

      Drosophila melanogaster SL-2 cells were treated for 8 days with ISWI (lane 1) or control (lane 2) dsRNAs. Total cell extracts (30 µg) were blotted with the acetylated ISWI (K753) polyclonal antibody, diluted 1/500.

       

    References for Anti-ISWI (acetyl K753) antibody (ab10748)

    This product has been referenced in:
    • 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) »
    • 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 . Read more (PubMed: 19995906) »

    See all 2 Publications for this product

    Product Wall

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step 0.1% Triton X-100, 0.1% BSA, 10% FCS, 0.1% sodium deoxycholate and 0.02% Thiomersal, an anti-fungal agent, in 1X PBS as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 24°C
    Sample Fruit fly (Drosophila melanogaster) Cell (Malpighian tubules, Salivary glands, Gut, etc)
    Specification Malpighian tubules, Salivary glands, Gut, etc
    Permeabilization Yes - 1% Triton X
    Fixative Paraformaldehyde
    Username

    Mr. Deo Prakash Deo Prakash

    Verified customer

    Submitted Jun 20 2013

    Thank you for your email. I am sorry that I have not been clear enough yesterday. To receive the reward points it is necessary to submit an Abreview over the System. You will find the link to fill in all the data and attach the image on the product pag...

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    Thank you very much for your Email and this valuable information. We appreciate the time that was spent on this experiments and putting all the data together a lot.

    In order to proceed in this case I like to ask you to submit an Abreview for A...

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    Thank you for your email. I am pleased to hear your customer has been working with the Fast-track antibody ab10748, directed against ISWI (acetyl K753).

    The customer would be eligible to receive Abpoints in return for their efforts which can b...

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    Thank you for your answer.

    Have you had the opportunity to test the antibody in Western blot as well? Indeed, as you have a positive control, I can recommend strongly to test the antibody in Western blot to see whether the antibody has retain...

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    Thank you for contacting us. I am sorry to hear that the results have not been as expected so far.

    Unfortunately, we are not able to test this antibody hereas it is difficult to get the positive controls. This is also why this antibody is sti...

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    Thank you for your enquiry and your interest.

    As the datasheet indicates this antibody recognizes Drosophila melanogaster ISWI (acetyl K753). However, it is not a fully characterized antibody, it is a Fast track antibody. It has been affinity...

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    Thank you for your enquiry. I am pleased that you have expressed an interest in this product. This is an in house product and the images are derived from a collaborator. We do not as yet possess any specific published references relating to this produc...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"