Recombinant
RabMAb

Recombinant Anti-HMGA1 antibody [EPR7839] - BSA and Azide free (ab226112)

Overview

  • Product name

    Anti-HMGA1 antibody [EPR7839] - BSA and Azide free
    See all HMGA1 primary antibodies
  • Description

    Rabbit monoclonal [EPR7839] to HMGA1 - BSA and Azide free
  • 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 (the amino acid sequence is considered to be commercially sensitive) 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 cell lysates, 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

    Ab226112 is the carrier-free version of ab129153. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab226112 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab226112 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 Use at an assay dependent concentration. Detects a band of approximately 17 kDa (predicted molecular weight: 12 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration.

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

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129153).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129153).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129153).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129153).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129153).

  • This IHC data was generated using the same anti-HMGA1 antibody clone, EPR7839, in a different buffer formulation (cat# ab129153).

    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.

  • This ICC data was generated using the same anti-HMGA1 antibody clone, EPR7839, in a different buffer formulation (cat# ab129153).

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

References

This product has been referenced in:

  • Hu J & Shen W microRNA-196a attenuates ischemic brain injury in rats by directly targeting high mobility group A1. Exp Ther Med 17:1579-1586 (2019). Read more (PubMed: 30783424) »
See 1 Publication for this product

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