Anti-HMGA2 antibody [EPR23215-46]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-HMGA2 antibody [EPR23215-46] (AB246513)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell) cells labelling HMGA2 with ab246513 at 1/600 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-HMGA2 antibody [EPR23215-46] (AB246513)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 cells labelling HMGA2 with ab246513 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing nuclear staining in HepG2 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 (2 ug/ml) dilution.

• IP

Unknown

Immunoprecipitation - Anti-HMGA2 antibody [EPR23215-46] (AB246513)

HMGA2 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate with ab246513 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab246513 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate 10 ug

Lane 2 : 246513 IP in HepG2 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab246513 in HepG2 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 minutes

All lanes:

Immunoprecipitation - Anti-HMGA2 antibody [EPR23215-46] (ab246513)

Predicted band size: 12 kDa

false

• WB

Lab

Western blot - Anti-HMGA2 antibody [EPR23215-46] (AB246513)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile and molecular weight observed is consistent with what has been described in the literature (PMID : 28765279)

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-HMGA2 antibody [EPR23215-46] (ab246513) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 40 µg

Lane 2:

NCI-H1299 (human lung carcinoma epithelial cell), whole cell lysate at 40 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 12 kDa

Observed band size: 18 kDa

false