Overview

  • Product nameAnti-HLTF antibody - ChIP GradeSee all HLTF primary antibodies ...
  • Description
    Rabbit polyclonal to HLTF - ChIP Grade
  • Tested applicationsChIP, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Rabbit, Guinea pig, Cow, Dog, Pig, Chimpanzee, Rhesus monkey, Gorilla, Orangutan, Platypus (Ornithorhynchus anatinus)
  • Immunogen

    Synthetic peptide.

  • EpitopeThe epitope recognized by antibody maps to a region between residues 600 and 700 of human SMARC A3 using the number given in entry NP_003062.2 (GeneID 6596)

Properties

Applications

Our Abpromise guarantee covers the use of ab17984 in the following tested applications.

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

Application Abreviews Notes
ChIP Use at an assay dependent concentration.
WB 1/3000 - 1/10000. Detects a band of approximately 125 kDa (predicted molecular weight: 114 kDa).
IP Use a concentration of 5 µg/ml.

Target

  • FunctionHas both helicasee and E3 ubiquitin ligase activities. Possesses intrinsic ATP-dependent nucleosome-remodeling activity; This activity may be required for transcriptional activation or repression of specific target promoters (By similarity). These may include the SERPINE1 and HIV-1 promoters and the SV40 enhancer, to which this protein can bind directly. Plays a role in error-free postreplication repair (PRR) of damaged DNA and maintains genomic stability through acting as a ubiquitin ligase for 'Lys-63'-linked polyubiquitination of chromatin-bound PCNA.
  • Tissue specificityExpressed in brain, heart, kidney, liver, lung, pancreas, placenta and skeletal muscle.
  • PathwayProtein modification; protein ubiquitination.
  • Sequence similaritiesBelongs to the SNF2/RAD54 helicase family. RAD16 subfamily.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.
    Contains 1 RING-type zinc finger.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • DNA-binding protein/plasminogen activator inhibitor 1 regulator antibody
    • Helicase like transcription factor antibody
    • Helicase-like transcription factor antibody
    • HIP116 antibody
    • HIP116A antibody
    • HLTF 1 antibody
    • HLTF antibody
    • HLTF_HUMAN antibody
    • HLTF1 antibody
    • p113 antibody
    • RING finger protein 80 antibody
    • RNF80 antibody
    • SMARC A3 antibody
    • SMARCA 3 antibody
    • SMARCA3 antibody
    • SNF2-like 3 antibody
    • SNF2L3 antibody
    • Sucrose nonfermenting protein 2 like 3 antibody
    • Sucrose nonfermenting protein 2-like 3 antibody
    • SWI/SNF related matrix associated actin dependent regulator of chromatin subfamily A member 3 antibody
    • SWI/SNF-related matrix-associated actin-dependent regulator of chromatin a3 antibody
    • SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 3 antibody
    • ZBU 1 antibody
    • ZBU1 antibody
    see all

Anti-HLTF antibody - ChIP Grade images

  • SMARCA3 was immunoprecipitated using 0.5mg HeLa whole cell extract, 5µg of Rabbit polyclonal to HLTF 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 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab17984.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 113kDa; SMARCA3, non specific bands - 50kDa: We are unsure as to the identity of this extra band.



  • Predicted band size : 114 kDa

    Detection of SMARCA3 by western blot and immunoprecipitation.
    Lane 1:  20 µg of HeLa nuclear extract
    Lane 2:  50 µg of HeLa nuclear extract
    LAne 3:  1 mg of HeLa nuclear extract immunoprecipitated by anti-SMARCA3 (ab17984), used at 4 µg/mg of lysate.
    Anti-SMARCA3 (ab17984) was used for western blot analysis at 1/5,000.
    Detection:  chemiluminescence with an exposure time of 4 minutes.

     

     

     

     

    Detection of SMARCA3 by western blot and immunoprecipitation.
    Lane 1:  20 µg of HeLa nuclear extract
    Lane 2:  50 µg of HeLa nuclear extract
    LAne 3:  1 mg of HeLa nuclear extract immunoprecipitated by anti-SMARCA3 (ab17984), used at 4 µg/mg of lysate.
    Anti-SMARCA3 (ab179

  • ab17984 in ChIP of rabbit endometrium nuclear cell lysate. In vivo binding of SMARCA3 (RUSH) to DNA and Egr-1/c-Rel in the rabbit RUSH promoter was confirmed by PCR amplification of products from ChIP assays.

    Lane 1 FX174/HaeIII size markers.

    Lane 2 Single 166-bp amplicons of the SMARCA3 (RUSH) site resulting from ChIP with antibodies to SMARCA3.

    Lanes 3 Negative ChIP control (IgG)

    Lane 4 PCR control (water)

    Lane 5 Single 153-bp amplicons of the Sp1(A) site resulting from ChIP with antibodies to SMARCA3. The SMARCA3 (RUSH) site was identified through antibody-SMARCA3-DNA binding. The Sp1(A) site is an Egr-1/Sp1/3/MAZ/MZF1/c-Rel composite binding site that was identified through antibody-SMARCA3-Egr-1/c-Rel-DNA binding.

    Cross-linking agent: disuccinimydyl glutarate and formaldehyde (0.75%)

    See Abreview

References for Anti-HLTF antibody - ChIP Grade (ab17984)

This product has been referenced in:
  • Helmer RA  et al. Helicase-like transcription factor (hltf) regulates g2/m transition, wt1/gata4/hif-1a cardiac transcription networks, and collagen biogenesis. PLoS One 8:e80461 (2013). Mouse . Read more (PubMed: 24278285) »
  • Helmer RA  et al. Prolactin-induced Jak2 phosphorylation of RUSH: a key element in Jak/RUSH signaling. Mol Cell Endocrinol 325:143-9 (2010). Read more (PubMed: 20562009) »

See all 3 Publications for this product

Product Wall

Thank you for your enquiry and interest in our products. As our Western blot image indicates on the product datasheet, ab17984 has been tested and characterized for Western blot on HeLa nuclear extracts so this type of lysate can be applied as posit...

Read More
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% TritonX100 in PBS
Username

Dr. Kirk McManus

Verified customer

Submitted Jun 15 2010

Application ChIP
Sample Rabbit Cell lysate - nuclear (endometrium)
Specification endometrium
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Detection step Semiquantitative PCR
Positive control PCR amplification of known postive sites.
Negative control No antibody for the ChIP, which became a negative control in the PCR reaction, plus a water blank for the PCR reaction.
Username

Prof. Beverly Chilton

Verified customer

Submitted Aug 16 2006

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