Anti-ESM1 antibody [RM1338]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ESM1 antibody [RM1338] (AB323845)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HUVEC (human umbilical vein endothelial cell) cells labelling ESM1 with ab323845 at 1/500 (0.998 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ab150081 1000 2ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in HUVEC cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : HL-60 (PMID : 8702785).
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/ab195889 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/ab150081 1000 2ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-ESM1 antibody [RM1338] (AB323845)

ESM1 was immunoprecipitated from 0.35 mg HUVEC (human umbilical vein endothelial cell) whole cell lysate with ab323845 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323845 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-ESM1 antibody [RM1338] (ab323845) at 1/1000 dilution

Lane 1:

HUVEC (human umbilical vein endothelial cell) whole cell lysate at 10 µg

Lane 2:

ab323845 at 1/30 IP in HUVEC (human umbilical vein endothelial cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab323845 in HUVEC 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

Observed band size: 20 kDa

false

Exposure time: 15s

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ESM1 antibody [RM1338] (AB323845)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized EL4 (mouse lymphoma T lymphocyte) cells labelling ESM1 with ab323845 at 1/500 (0.998 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ab150081 1000 2ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in EL4 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : Neuro-2a.
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/ab195889 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/ab150081 1000 2ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-ESM1 antibody [RM1338] (AB323845)

ESM1 was immunoprecipitated from 0.35 mg EL4 (mouse lymphoma T lymphocyte) whole cell lysate with ab323845 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323845 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-ESM1 antibody [RM1338] (ab323845) at 1/1000 dilution

Lane 1:

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

Lane 2:

ab323845 at 1/30 IP in EL4 (mouse lymphoma T lymphocyte) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab323845 in EL4 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

Observed band size: 14 kDa,20 kDa

false

Exposure time: 32s

• WB

Supplier Data

Western blot - Anti-ESM1 antibody [RM1338] (AB323845)

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

Negative control : A549, HL-60 (PMID : 8702785)

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

All lanes:

Western blot - Anti-ESM1 antibody [RM1338] (ab323845) at 1/1000 dilution

Lane 1:

HUVEC (human umbilical vein endothelial cell) whole cell lysate at 40 µg

Lane 2:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 3:

HL-60 (human acute promyelocytic leukemia promyeloblast) 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 1/100000 dilution

Observed band size: 20 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-ESM1 antibody [RM1338] (AB323845)

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

Negative control : Neuro-2a.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 8702785).

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

All lanes:

Western blot - Anti-ESM1 antibody [RM1338] (ab323845) at 1/1000 dilution

Lane 1:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 40 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) 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 1/100000 dilution

Observed band size: 14 kDa,20 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-ESM1 antibody [RM1338] (AB323845)

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

Negative control : skeletal muscle.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 8702785).

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

All lanes:

Western blot - Anti-ESM1 antibody [RM1338] (ab323845) at 1/1000 dilution

Lane 1:

Mouse pituitary tissue lysate at 40 µg

Lane 2:

Mouse skeletal muscle tissue 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 1/100000 dilution

Observed band size: 14 kDa,20 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-ESM1 antibody [RM1338] (AB323845)

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

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

In Western blot, Anti-6X His tag^® antibody [AD1.1.10] (ab15149) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-ESM1 antibody [RM1338] (ab323845) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 10 µg

Lane 2:

293T cells transfected with a human ESM1 expression vector containing a myc-His-tag®, whole cell lysate at 10 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 1/100000 dilution

Observed band size: 20 kDa,36 kDa

false

Exposure time: 26s