Anti-HMGA1 antibody [RM1061]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1 antibody [RM1061] (AB315350)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling HMGA1 with ab315350 at 1/2000 (0.251 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on human testis in (A) ab315350 and (B) ab4078.
The image on (A) ab315350 is applied with Anti-HMGA1 antibody at 1/2000 dilution and (B) ab4078 is applied with Anti-HMGA1 antibody at 1/500 dilution.
The section was incubated with ab315350 or ab4078 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-HMGA1 antibody [RM1061] (AB315350)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NCCIT (human pluripotent embryonic carcinoma epithelial cell) cells labelling HMGA1 with ab315350 at 1/2000 (1.004 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing nuclear staining in NCCIT cell line.
Low expression：MCF7 (PMID : 12640109).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1 antibody [RM1061] (AB315350)

Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissue labeling HMGA1 with ab315350 at 1/2000 (0.251 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on human ovarian cancer.
The section was incubated with ab315350 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-HMGA1 antibody [RM1061] (AB315350)

HMGA1 was immunoprecipitated from 0.35 mg BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate with ab315350 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315350 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab315350 IP in BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab315350 in BxPC-3 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-HMGA1 antibody [RM1061] (ab315350) at 1/30 dilution

All lanes:

BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate

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

false

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1 antibody [RM1061] (AB315350)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling HMGA1 with ab315350 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat testis.
The section was incubated with ab315350 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1 antibody [RM1061] (AB315350)

Immunohistochemical analysis of paraffin-embedded Mouse lung carcinoma tissue labeling HMGA1 with ab315350 at 1/5000 (0.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung carcinoma.
The section was incubated with ab315350 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• WB

Supplier Data

Western blot - Anti-HMGA1 antibody [RM1061] (AB315350)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-HMGA1 antibody [RM1061] (ab315350) at 1/1000 dilution

Lane 1:

BxPC-3 (human pancreas adenocarcinoma epithelial cell) nuclear fraction at 20 µg

Lane 2:

BxPC-3 (human pancreas adenocarcinoma epithelial cell) cytoplasmic fraction at 20 µg

Lane 3:

BxPC-3 (human pancreas adenocarcinoma epithelial cell) membrane fraction 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

Observed band size: 17 kDa,36 kDa,15 kDa,125 kDa

false

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-HMGA1 antibody [RM1061] (AB315350)

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

Negative control : MCF7 (PMID : 12640109).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-HMGA1 antibody [RM1061] (ab315350) at 1/1000 dilution

Lane 1:

BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 40 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

Observed band size: 17 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-HMGA1 antibody [RM1061] (AB315350)

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

Negative control : 293T (PMID : 27445976), MCF7 (PMID : 12640109).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1 : 180 seconds Lane 2-6 : 81 seconds

All lanes:

Western blot - Anti-HMGA1 antibody [RM1061] (ab315350) at 1/1000 dilution

Lane 1:

NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HCT-15 (human colon epithelial cell) whole cell lysate at 20 µg

Lane 5:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 17 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-HMGA1 antibody [RM1061] (AB315350)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1 : 103 seconds Lane 2 : 59 seconds Lane 3 : 180 seconds

All lanes:

Western blot - Anti-HMGA1 antibody [RM1061] (ab315350) at 1/1000 dilution

Lane 1:

EL4 (mouse lymphoma T lymphocyte) nuclear fraction at 48 µg

Lane 2:

C2C12 (mouse myoblast) nuclear fraction at 48 µg

Lane 3:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 17 kDa,36 kDa,15 kDa

false