Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078)

Overview

  • Product nameAnti-HMGA1a / HMGA1b antibody - ChIP Grade
    See all HMGA1a / HMGA1b primary antibodies
  • Description
    Rabbit polyclonal to HMGA1a / HMGA1b - ChIP Grade
  • SpecificityThis antibody detects, by Western blot on human GFP fusion proteins in cell lysate, both HMGA1a and the splice variant HMGA1b, but does not detect HMGA2 (see review and Western blot picture). The antibody has not yet been successfully used to detect endogenous protein.
  • Tested applicationsSuitable for: WB, ChIP, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human, Zebrafish
    Predicted to work with: Rat, Chicken, Neurospora crassa , Chinese Hamster
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human HMGA1a/ HMGA1b.

    (Peptide available as ab20073.)

  • Positive control
    • This antibody gave a positive result in human placenta FFPE tissue sections

Properties

Applications

Our Abpromise guarantee covers the use of ab4078 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
WB
ChIP
IHC-P
ICC/IF
  • Application notesChIP: Use at an assay dependent dilution. See Abreview submitted on 15 December 2009.
    ICC/IF: 1/250 - 1/500. Although this antibody gives a specific signal in Immunofluoresence, we have had suggestions that strong background signals are seen. See review by Robert Hock and IF picture below.
    IHC-P: Use at an assay dependent dilution.
    WB: 1/1000. Detects a band of approximately 11 kDa (predicted molecular weight: 11.5 kDa).

    Not tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • FunctionHMG-I/Y bind preferentially to the minor groove of A+T rich regions in double stranded DNA. It is suggested that these proteins could function in nucleosome phasing and in the 3'-end processing of mRNA transcripts. They are also involved in the transcription regulation of genes containing, or in close proximity to A+T-rich regions.
    • Involvement in diseaseNote=A chromosomal aberration involving HMGA1 is found in pulmonary chondroid hamartoma. Translocation t(6;14)(p21;q23-24) with RAD51L1.
    • Sequence similaritiesBelongs to the HMGA family.
      Contains 3 A.T hook DNA-binding domains.
    • Post-translational
      modifications
      Constitutively phosphorylated on two or three sites. Phosphorylated upon DNA damage, probably by ATM or ATR. Hyperphosphorylated at early stages of apoptosis, followed by dephosphorylation and methylation, which coincides with chromatin condensation. Isoform HMG-Y can be phosphorylated by HIPK2.
      HMG-Y is not methylated.
      Methylation at Arg-58 is mutually exclusive with methylation at Arg-60.
    • Cellular localizationNucleus. Chromosome.
    • Information by UniProt
    • Database links
    • Alternative names
      • High mobility group AT hook 1 antibody
      • High mobility group AT-hook protein 1 antibody
      • High mobility group protein A1 antibody
      • High mobility group protein HMG-I/HMG-Y antibody
      • High mobility group protein R antibody
      • HMG-I(Y) antibody
      • HMGA 1a antibody
      • HMGA 1b antibody
      • HMGA1 antibody
      • HMGA1_HUMAN antibody
      • HMGI antibody
      • HMGI/Y antibody
      • HMGR antibody
      • HMGY antibody
      see all

    Anti-HMGA1a / HMGA1b antibody - ChIP Grade images

    • Immunofluoresence on MCF-7 cells transfected with human HMGA1a-GFP using ab4078.

      A: Hoechst stain
      B: Phasecontrast
      C: TexasRed HMG-I/HMG-Y (ab4078)
      D: HMGA1a-GFP
      E: Merge

    • Lanes 1 - 4 : Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078) at 1/1000 dilution
      Lanes 5 - 8 : Anti-GFP

      Lane 1 : Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA1a GFP fusion proteins.
      Lane 2 : Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA1b-GFP fusion proteins
      Lane 3 : Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA2-GFP fusion proteins
      Lane 4 : hole cell lysate from HepG2 cells transfected with plasmids coding for NLS-GFP fusion proteins
      Lane 5 : Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA1a-GFP fusion proteins
      Lane 6 : Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA1b-GFP fusion proteins
      Lane 7 : Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA2-GFP fusion proteins
      Lane 8 : Whole cell lysate from HepG2 cells transfected with plasmids coding for NLS-GFP fusion proteins

      Secondary
      Anti-rabbit HRP at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size : 11.5 kDa

      This image is courtesy of Monika Harrer

      Western blot using ab4078.

      Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA1a-, hHMGA1b-, hHMGA2- or NLS- (nuclear localisation signal) GFP fusion proteins.

      ab4078 was diluted 1/1000 in 5% milk/TBS and incubated for 1 hr at room temperature. Secondary HRP anti-rabbit antibody was diluted 1/10000 in 5% milk/TBS and incubated for 1 hr at room temperature.

      Lane 1: hHMGA1a-GFP (ab4078)
      Lane 2: hHMGA1b-GFP (ab4078)
      Lane 3: hHMGA2-GFP (ab4078)
      Lane 4: NLS-GFP (ab4078)
      Lane 5: hHMGA1a-GFP (anti-GFP)
      Lane 6: hHMGA1b-GFP (anti-GFP)
      Lane 7: hHMGA2-GFP (anti-GFP)
      Lane 8: NLS-GFP (anti-GFP)

      The detected size is that expected for the fusion of an 11kD and 26kD protein (GFP).

    • Chromatin was prepared from the Human Embryonic Stem Cells. The cross-linking (X-ChiP) technique was used; crosslinking was performed for 15 minutes in formaldehyde. The primary antibody was diluted 1/100 in IP dilution buffer and incubated with the sample for 12 hours at 4°C. Histone H3 was used as the positive control, whilst normal rabbit IgG was used as the negative control. The immunoprecipitated DNA was quantified by real time PCR.
    • IHC image of ab4078 staining in human placenta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab4078, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    References for Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078)

    This product has been referenced in:
    • Arce-Cerezo A  et al. HMGA1 overexpression in adipose tissue impairs adipogenesis and prevents diet-induced obesity and insulin resistance. Sci Rep 5:14487 (2015). WB ; Mouse . Read more (PubMed: 26411793) »
    • Qu Y  et al. Overexpression of high mobility group A1 protein in human uveal melanomas: implication for prognosis. PLoS One 8:e68724 (2013). IHC ; Human . Read more (PubMed: 23935884) »

    See all 8 Publications for this product

    Product Wall

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement (from a different lot) with the order number 949551. To check the status of t...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Zebrafish Tissue lysate - whole (1.5 zebrafish embryo (74hpf)/lane)
    Specification 1.5 zebrafish embryo (74hpf)/lane
    Gel Running Conditions Reduced Denaturing (13%)
    Blocking step Milk as blocking agent for 1 hour(s) and 20 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Dr. Jan Brocher

    Verified customer

    Submitted Apr 20 2010

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application ChIP
    Sample Human Cell lysate - nuclear (Human embryonic stem cells)
    Specification Human embryonic stem cells
    Type Cross-linking (X-ChIP)
    Duration of cross-linking step: 15 minute(s) and 0 second(s)
    Specification of the cross-linking agent: formaldehyde
    Detection step Real-time PCR
    Positive control Histone H3 antibody from Abcam
    Negative control Rabbit IgG
    Username

    Abcam user community

    Verified customer

    Submitted Dec 15 2009

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