Product nameAnti-HMGA1a / HMGA1b antibody - ChIP Grade
See all HMGA1a / HMGA1b primary antibodies
DescriptionRabbit 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 reactivityReacts with: Mouse, Human, Zebrafish
Predicted to work with: Rat, Chicken, Neurospora crassa, Chinese hamster
- This antibody gave a positive result in human placenta FFPE tissue sections
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
ChIP Related Products
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.
|WB||1/1000. Detects a band of approximately 11 kDa (predicted molecular weight: 11.5 kDa).|
|ChIP||Use at an assay dependent concentration.
See Abreview submitted on 15 December 2009.
|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.
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.
modificationsConstitutively 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
- High mobility group AT hook 1 antibody
- High mobility group AT-hook protein 1 antibody
- High mobility group protein A1 antibody
Immunofluoresence on MCF-7 cells transfected with human HMGA1a-GFP using ab4078.
A: Hoechst stain
C: TexasRed HMG-I/HMG-Y (ab4078)
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
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).
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.
This product has been referenced in:
- Zhang H et al. Mandatory role of HMGA1 in human airway epithelial normal differentiation and post-injury regeneration. Oncotarget 9:14324-14337 (2018). Read more (PubMed: 29581847) »
- Parry AJ et al. NOTCH-mediated non-cell autonomous regulation of chromatin structure during senescence. Nat Commun 9:1840 (2018). Read more (PubMed: 29743479) »