Overview

  • Product name
    Anti-HMGA1 antibody [EPR7839]
    See all HMGA1 primary antibodies
  • Description
    Rabbit monoclonal [EPR7839] to HMGA1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human HMGA1 (N terminal). The exact sequence is proprietary.
    Database link: P17096

  • Positive control
    • WB: SK-OV-3, Caco-2, BxPC-3 and HepG2 whole cell lysate (ab7900), rat kidney and mouse brain tissue lysates. IHC-P: Human testis and transitional cell carcinoma of bladder tissues. ICC/IF: HeLa and BxPC-3 cells. Flow Cyt: HepG2 cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab129153 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 1/10000 - 1/50000. Detects a band of approximately 17 kDa (predicted molecular weight: 12 kDa).
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Flow Cyt 1/60.

For unpurified use at 1/100 - 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/500.

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
  • Alternative names
    • High mobility group (nonhistone chromosomal) protein isoforms I and Y antibody
    • High mobility group AT hook 1 antibody
    • High mobility group AT-hook protein 1 antibody
    • High mobility group protein 1 antibody
    • High mobility group protein A1 antibody
    • High mobility group protein HMG-I/HMG-Y antibody
    • High mobility group protein Ia antibody
    • High mobility group protein R antibody
    • HMG R antibody
    • HMG-I(Y) antibody
    • HMGA1 antibody
    • HMGA1_HUMAN antibody
    • HMGA1A antibody
    • HMGIY antibody
    • MGC12816 antibody
    • MGC4242 antibody
    • MGC4854 antibody
    • Nonhistone chromosomal high mobility group protein HMG I/HMG Y antibody
    see all

Images

  • All lanes : Anti-HMGA1 antibody [EPR7839] (ab129153) (purified)

    Lane 1 : HepG2 cell lysate
    Lane 2 : SK-OV-3 cell lysate
    Lane 3 : Caco-2 cell lysate
    Lane 4 : BxPC-3 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 12 kDa
    Observed band size: 17 kDa
    why is the actual band size different from the predicted?



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunocytochemistry/Immunofluorescence analysis of BxPC-3 cells labelling HMGA1 with unpurified ab129153 at 1/250 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human transitional cell carcinoma of bladder tissue labelling HMGA1 with purified ab129153 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

  • Anti-HMGA1 antibody [EPR7839] (ab129153) at 1/10000 dilution (purified) + Mouse brain tissue lysate at 10 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 12 kDa
    Observed band size: 17 kDa why is the actual band size different from the predicted?



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Anti-HMGA1 antibody [EPR7839] (ab129153) at 1/50000 dilution (purified) + Rat kidney tissue lysate at 10 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 12 kDa
    Observed band size: 17 kDa why is the actual band size different from the predicted?



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-HMGA1 antibody [EPR7839] (ab129153) at 1/10000 dilution (unpurified)

    Lane 1 : SK-OV-3 cell lysate
    Lane 2 : Caco-2 cell lysate
    Lane 3 : BxPC-3 cell lysate
    Lane 4 : HepG2 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size: 12 kDa
    Observed band size: 17 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling HMGA1 with unpurified ab129153 at 1/250.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling HMGA1 with purified ab129153 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

  • Flow cytometry analysis of HepG2 cells labelling HMGA1 with purified ab129153 at 1/60 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Overlay histogram showing HepG2 cells stained with unpurified ab129153 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab129153, 1/870 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:
  • Kong B  et al. microRNA-26a-5p affects myocardial injury induced by coronary microembolization by modulating HMGA1. J Cell Biochem N/A:N/A (2019). Read more (PubMed: 30652345) »
  • Zaatiti H  et al. Tumorigenic proteins upregulated in the MYCN-amplified IMR-32 human neuroblastoma cells promote proliferation and migration. Int J Oncol 52:787-803 (2018). Read more (PubMed: 29328367) »
See all 22 Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Retina)
Permeabilization
Yes - Triton X 100
Specification
Retina
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 07 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Zytomed Systems HRP Polymer Kit BLocking reagent as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 25°C
Antigen retrieval step
None
Sample
Human Tissue sections (Lung cancer)
Specification
Lung cancer
Permeabilization
No
Fixative
HOPE

Abcam user community

Verified customer

Submitted Mar 19 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Zytomed Systems Blocking HRP Polymer SystemReagent as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 25°C
Sample
Human Cell (paraffin-embedded human lung cancer tissue)
Specification
paraffin-embedded human lung cancer tissue
Permeabilization
No
Fixative
HOPE

Abcam user community

Verified customer

Submitted Mar 19 2014

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

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