Anti-HMGA1 antibody [EPR7839] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-HMGA1 antibody [EPR7839] - BSA and Azide free (AB226112)

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).

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-HMGA1 antibody [EPR7839] - BSA and Azide free (AB226112)

This data was developed using the same antibody clone in a different buffer formulation (ab129153).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 (human hepatocellular carcinoma epithelial cell) cells and 5 µg of ab129153 [EPR7839]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-HMGA1 antibody [EPR7839] - BSA and Azide free (AB226112)

This data was developed using the same antibody clone in a different buffer formulation (ab129153).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 (human hepatocellular carcinoma epithelial cell) cells and 5 µg of ab129153 [EPR7839]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-HMGA1 antibody [EPR7839] - BSA and Azide free (AB226112)

This data was developed using the same antibody clone in a different buffer formulation (ab129153).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 (human hepatocellular carcinoma epithelial cell) cells and 5 µg of ab129153 [EPR7839]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-HMGA1 antibody [EPR7839] - BSA and Azide free (AB226112)

Intracellular 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).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1 antibody [EPR7839] - BSA and Azide free (AB226112)

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.

• WB

Lab

Western blot - Anti-HMGA1 antibody [EPR7839] - BSA and Azide free (AB226112)

Blocking buffer and concentration : 5% NFDM/TBST

Diluting buffer and concentration : 5% NFDM /TBST

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

Lane 1:

Western blot - Anti-HMGA1 antibody [EPR7839] - BSA and Azide free (ab226112) at 1/10000 dilution

Lane 1:

Western blot - Anti-HMGA1 antibody [EPR7839] (<a href='/en-us/products/primary-antibodies/hmga1-antibody-epr7839-ab129153'>ab129153</a>) at 1/10000 dilution

All lanes:

SK-OV-3 (Human ovarian cancer epithelial cell) whole cell 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: 5s

• WB

Lab

Western blot - Anti-HMGA1 antibody [EPR7839] - BSA and Azide free (AB226112)

Blocking buffer and concentration : 5% NFDM/TBST Diluting buffer and concentration : 5% NFDM /TBST This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129153).

Lanes 1 - 3:

Western blot - Anti-HMGA1 antibody [EPR7839] - BSA and Azide free (ab226112) at 1/1000 dilution

Lanes 1 - 3:

Western blot - Anti-HMGA1 antibody [EPR7839] (<a href='/en-us/products/primary-antibodies/hmga1-antibody-epr7839-ab129153'>ab129153</a>) at 1/1000 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: 3s

• WB

Lab

Western blot - Anti-HMGA1 antibody [EPR7839] - BSA and Azide free (AB226112)

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

Lanes 1 - 4:

Western blot - Anti-HMGA1 antibody [EPR7839] - BSA and Azide free (ab226112) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-HMGA1 antibody [EPR7839] (<a href='/en-us/products/primary-antibodies/hmga1-antibody-epr7839-ab129153'>ab129153</a>) at 1/1000 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: 5s