Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Key features and details

  • Rabbit polyclonal to HMGA1a / HMGA1b - ChIP Grade
  • Suitable for: WB, IHC-P, ICC/IF
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-HMGA1a / HMGA1b antibody - ChIP Grade
    See all HMGA1a / HMGA1b primary antibodies

  • Description

    Rabbit polyclonal to HMGA1a / HMGA1b - ChIP Grade

  • Host species

    Rabbit

  • Specificity

    This 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 applications

    Suitable for: WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Chicken, Zebrafish, Neurospora crassa, Chinese hamster

  • Immunogen

    Synthetic peptide corresponding to Human HMGA1a/ HMGA1b aa 1-100 conjugated to keyhole limpet haemocyanin.
    (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 1/1000. Detects a band of approximately 11 kDa (predicted molecular weight: 11.5 kDa).
IHC-P Use at an assay dependent concentration.
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.

Target

  • Function

    HMG-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 disease

    Note=A chromosomal aberration involving HMGA1 is found in pulmonary chondroid hamartoma. Translocation t(6;14)(p21;q23-24) with RAD51L1.

  • Sequence similarities

    Belongs 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 localization

    Nucleus. Chromosome.
  • Information by UniProt

  • Database links

    see all

  • 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

Images

  • Western blot - Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078)
    Western blot - Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078)This image is courtesy of Monika Harrer
    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.
    Lanes 2 & 6 : Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA1b-GFP fusion proteins
    Lanes 3 & 7 : 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 8 : Whole cell lysate from HepG2 cells transfected with plasmids coding for NLS-GFP fusion proteins

    Secondary
    All lanes : Anti-rabbit HRP at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 11.5 kDa



    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078)
    Immunocytochemistry/ Immunofluorescence - Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078)This image is courtesy of Robert Hock

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078)
    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 (22)

Publishing research using ab4078? Please let us know so that we can cite the reference in this datasheet.

ab4078 has been referenced in 22 publications.

  • Teng K  et al. KIFC1 is activated by TCF-4 and promotes hepatocellular carcinoma pathogenesis by regulating HMGA1 transcriptional activity. J Exp Clin Cancer Res 38:329 (2019). PubMed: 31340839
  • Tolza C  et al. AP-1 Signaling by Fra-1 Directly Regulates HMGA1 Oncogene Transcription in Triple-Negative Breast Cancers. Mol Cancer Res 17:1999-2014 (2019). PubMed: 31300541
  • Patel ZH  et al. A plausibly causal functional lupus-associated risk variant in the STAT1-STAT4 locus. Hum Mol Genet 27:2392-2404 (2018). PubMed: 29912393
  • Zhang H  et al. Mandatory role of HMGA1 in human airway epithelial normal differentiation and post-injury regeneration. Oncotarget 9:14324-14337 (2018). PubMed: 29581847
  • Parry AJ  et al. NOTCH-mediated non-cell autonomous regulation of chromatin structure during senescence. Nat Commun 9:1840 (2018). PubMed: 29743479
View all Publications for this product

Images

  • Western blot - Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078)
    Western blot - Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078) This image is courtesy of Monika Harrer
    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.
    Lanes 2 & 6 : Whole cell lysate from HepG2 cells transfected with plasmids coding for hHMGA1b-GFP fusion proteins
    Lanes 3 & 7 : 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 8 : Whole cell lysate from HepG2 cells transfected with plasmids coding for NLS-GFP fusion proteins

    Secondary
    All lanes : Anti-rabbit HRP at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 11.5 kDa



    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078)
    Immunocytochemistry/ Immunofluorescence - Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078) This image is courtesy of Robert Hock

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1a / HMGA1b antibody - ChIP Grade (ab4078)
    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.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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