Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (AB255277)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling HMGA1 with ab252930 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in germ cells of human testis (PMID : 17935122) is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252930).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (AB255277)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized MCF7 (Human breast adenocarcinoma epithelial cell, Left) / NCCIT (Human pluripotent embryonic carcinoma epithelial cell, Right) cell lines labeling HMGA1 with ab252930 at 1/600 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody. Weak expression control : MCF7 (PMID : 12640109).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252930).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (AB255277)

Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue labeling HMGA1 with ab252930 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human pancreas cancer (PMID : 18473350) is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252930).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (AB255277)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NCCIT (human pluripotent embryonic carcinoma epithelial cell) cells labeling HMGA1 with ab252930 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in NCCIT cell line. Low expression cell line : MCF7?PMID : 12640109). The nuclear counter stain is DAPI (blue). Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).

The negative control is the secondary antibody only.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252930).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (AB255277)

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling HMGA1 with ab252930 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human lung cancer (PMID : 25344216) is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252930).

• IP

Supplier Data

Immunoprecipitation - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (AB255277)

HMGA1 was immunoprecipitated from 0.35mg of EL4 (mouse lymphoma T lymphocyte) whole cell lysate with ab252930 at 1/30 dilution.

Blocking and dilution 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 (ab252930).

All lanes:

Immunoprecipitation - Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade (<a href='/en-us/products/primary-antibodies/hmga1-antibody-epr22421-101-chip-grade-ab252930'>ab252930</a>) at 1/1000 dilution

Lane 1:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/hmga1-antibody-epr22421-101-chip-grade-ab252930'>ab252930</a> IP in EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/hmga1-antibody-epr22421-101-chip-grade-ab252930'>ab252930</a> in EL4 (human epidermoid carcinoma) whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Predicted band size: 17 kDa

Observed band size: 12 kDa

false

Exposure time: 180s

• ChIP

Supplier Data

ChIP - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (AB255277)

Chromatin was prepared from EL4 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min.

The ChIP was performed with 25 μg of chromatin, 5 μg of ab252930 (red) or 5 μg of rabbit normal IgG (gray) was added to 20 μl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (sybr green approach).

Primers and probes are located in the first kb of the transcribed region.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252930).

• IP

Unknown

Immunoprecipitation - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (AB255277)

HMGA1 was immunoprecipitated from 0.35 mg HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate with ab252930 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab252930 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.

Lane 1 : HCT 116 whole cell lysate 10 μg (Input).
Lane 2 : ab252930 IP in HCT 116 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab252930 in HCT 116 whole cell lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.
Exposure time : 40 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252930).

All lanes:

Immunoprecipitation - Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade (<a href='/en-us/products/primary-antibodies/hmga1-antibody-epr22421-101-chip-grade-ab252930'>ab252930</a>)

Predicted band size: 12 kDa

Observed band size: 17 kDa

false

• WB

Lab

Western blot - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (AB255277)

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 (ab252930).

Lanes 1 - 3:

Western blot - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277) at 1/1000 dilution

Lanes 1 - 3:

Western blot - Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade (<a href='/en-us/products/primary-antibodies/hmga1-antibody-epr22421-101-chip-grade-ab252930'>ab252930</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: 10s

• WB

Lab

Western blot - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (AB255277)

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 (ab252930).

Lanes 1 - 4:

Western blot - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade (<a href='/en-us/products/primary-antibodies/hmga1-antibody-epr22421-101-chip-grade-ab252930'>ab252930</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: 40s