HMGA2 Recombinant Rabbit Monoclonal Antibody [PSH05-53]
cat.: HA722295
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IF-Cell, IHC-P, FC, IF-Tissue
Clonality: Monoclonal
Clone number: PSH05-53
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 12 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human HMGA2 aa 70-120.
Positive control: HepG2 cell lysate, MCF7 cell lysate, NIH/3T3 cell lysate, NIH/3T3, human colon cancer tissue, mouse colon tissue.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IF-Tissue

1:1,000
1:1,000
1:8,000
1:1,000
1:200-1:1,000
Uniprot #: SwissProt: P52926 Human | P52927 Mouse
Alternative names: HMGA 2 BABL High mobility group (nonhistone chromosomal) protein isoform I C High mobility group (nonhistone chromosomal) protein isoform IC High Mobility Group AT hook 2 High Mobility Group AT hook protein 2 High mobility group AT-hook protein 2 High mobility group protein HMGI C High mobility group protein HMGI-C High mobility group protein HMGIC High mobility group protein I, isoform C HMGA2 HMGA2_HUMAN HMGI C HMGIC LIPO Non histone chromosomal architectural protein HMGI C Non histone chromosomal architectural protein HMGIC Pygmy STQTL9
Images
HA722295_1.jpg Fig1: Western blot analysis of HMGA2 on different lysates with Rabbit anti-HMGA2 antibody (HA722295) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: MCF7 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: NIH/3T3 cell lysate (no heat)

Notice: no heat means the lysate is not boiled.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 12 kDa
Observed band size: 18 kDa

Exposure time: 10 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722295) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA722295_2.jpg Fig2: Immunocytochemistry analysis of NIH/3T3 cells labeling HMGA2 with Rabbit anti-HMGA2 antibody (HA722295) at 1/2,500 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HMGA2 antibody (HA722295) at 1/2,500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA722295_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-HMGA2 antibody (HA722295) at 1/8,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722295) at 1/8,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA722295_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-HMGA2 antibody (HA722295) at 1/8,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722295) at 1/8,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA722295_5.jpg Fig5: Flow cytometric analysis of NIH/3T3 cells labeling HMGA2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722295, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.