Anti-Acid sphingomyelinase antibody [EPR28613-49] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Acid sphingomyelinase antibody [EPR28613-49] - BSA and Azide free (AB315811)

This data was developed using ab315810, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling Acid sphingomyelinase with ab315810 at 1/50 (9.8 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 increased cytoplasmic staining in RAW 264.7 cells treated with Phorbol-12-myristate-13-acetate (80nM) for 24 hours.
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 (Red). 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.

• WB

Supplier Data

Western blot - Anti-Acid sphingomyelinase antibody [EPR28613-49] - BSA and Azide free (AB315811)

This data was developed using ab315810, the same antibody clone in a different buffer formulation.

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

Low expression : lung, MOLT-4

The molecular weight observed is consistent with what has been described in the literature (PMID : 20807762).

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

All lanes:

Western blot - Anti-Acid sphingomyelinase antibody [EPR28613-49] (<a href='/en-us/products/primary-antibodies/acid-sphingomyelinase-antibody-epr28613-49-ab315810'>ab315810</a>) at 1/1000 dilution

Lane 1:

Rat liver tissue lysate at 20 µg

Lane 2:

Rat lung tissue lysate at 20 µg

Lane 3:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate 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 1/100000 dilution

Observed band size: 75 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Acid sphingomyelinase antibody [EPR28613-49] - BSA and Azide free (AB315811)

This data was developed using ab315810, the same antibody clone in a different buffer formulation.

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

Low expression : lung

The molecular weight observed is consistent with what has been described in the literature (PMID : 20807762).

Acid sphingomyelinase expression can be induced by PMA treatment(PMID : 10224156).

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

All lanes:

Western blot - Anti-Acid sphingomyelinase antibody [EPR28613-49] (<a href='/en-us/products/primary-antibodies/acid-sphingomyelinase-antibody-epr28613-49-ab315810'>ab315810</a>) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse lung tissue lysate at 20 µg

Lane 4:

Untreated RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 5:

RAW 264.7 treated with 100nM Phorbol-12-myristate-13-acetate (PMA) for 24 hours, whole cell lysate 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 1/100000 dilution

Observed band size: 75 kDa,36 kDa

false

Exposure time: 81s