Anti-HMGA1 antibody [EPR16649] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-HMGA1 antibody [EPR16649] - BSA and Azide free (AB251363)

This data was developed using ab202070, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized BxPC-3 (Human pancreas adenocarcinoma cells) cells labeling HMGA1 with ab202070 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on BxPC-3 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

The negative controls are as follows :
-ve control 1 : ab202070 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-HMGA1 antibody [EPR16649] - BSA and Azide free (AB251363)

This data was developed using ab202070, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling HMGA1 with ab202070 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-HMGA1 antibody [EPR16649] - BSA and Azide free (AB251363)

Blocking buffer and concentration : 5% NFDM/TBST Diluting buffer and concentration : 5% NFDM/TBST We recommend using 1% SDS Hot lysis prepare method to get desired Western Blot results. This data was developed using ab202070, the same antibody clone in a different buffer formulation.

Lanes 1 - 4:

Western blot - Anti-HMGA1 antibody [EPR16649] - BSA and Azide free (ab251363) at 1/10000 dilution

Lanes 1 - 4:

Western blot - Anti-HMGA1 antibody [EPR16649] (<a href='/en-us/products/primary-antibodies/hmga1-antibody-epr16649-ab202070'>ab202070</a>) at 1/10000 dilution

Lane 1:

HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method at 20 µg

Lane 2:

HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate prepared in 1% SDS Hot lysis method at 20 µg

Lane 3:

MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method at 20 µg

Lane 4:

MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate prepared in 1%SDS Hot lysis method at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 12 kDa

Observed band size: 17 kDa

false

Exposure time: 60s

• WB

Lab

Western blot - Anti-HMGA1 antibody [EPR16649] - BSA and Azide free (AB251363)

Blocking buffer and concentration : 5% NFDM/TBST Diluting buffer and concentration : 5% NFDM/TBST This data was developed using ab202070, the same antibody clone in a different buffer formulation.

Lanes 1 - 3:

Western blot - Anti-HMGA1 antibody [EPR16649] - BSA and Azide free (ab251363) at 1/10000 dilution

Lanes 1 - 3:

Western blot - Anti-HMGA1 antibody [EPR16649] (<a href='/en-us/products/primary-antibodies/hmga1-antibody-epr16649-ab202070'>ab202070</a>) at 1/10000 dilution

Lane 1:

MDA-MB-231 (Human breast adenocarcinoma epithelial cell) nuclear fraction lysate at 15 µg

Lane 2:

MDA-MB-231 (Human breast adenocarcinoma epithelial cell) cytoplasm fraction lysate at 15 µg

Lane 3:

MDA-MB-231 (Human breast adenocarcinoma epithelial cell) membrane fraction lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 12 kDa

Observed band size: 17 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-HMGA1 antibody [EPR16649] - BSA and Azide free (AB251363)

This data was developed using ab202070, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-HMGA1 antibody [EPR16649] (<a href='/en-us/products/primary-antibodies/hmga1-antibody-epr16649-ab202070'>ab202070</a>) at 1/10000 dilution

Lane 1:

NCCIT (Human pluripotent embryonal carcinoma) whole cell lysate at 10 µg

Lane 2:

BxPC-3 (Human pancreas adenocarcinoma cells) whole cell lysate at 10 µg

Lane 3:

HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 10 µg

Lane 4:

SK-OV-3 (Human ovarian cancer cell line) whole cell lysate at 10 µg

Lane 5:

Caco-2 (Human colorectal adenocarcinoma cells) whole cell lysate at 10 µg

Lane 6:

Human testis tissue lysate at 10 µg

Lane 7:

Human lung tissue lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 12 kDa

Observed band size: 18 kDa

false