Anti-ACSS2 antibody [EPR27496-4] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-ACSS2 antibody [EPR27496-4] - BSA and Azide free (AB314491)

This data was developed using ab314490, the same antibody clone in a different buffer formulation. ACSS2 was immunoprecipitated from 0.35 mg HAP1 (human chronic myelogenous leukemia near-haploid cell line) whole cell lysate with ab314490 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314490 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HAP1 (human chronic myelogenous leukemia near-haploid cell line) whole cell lysate Lane 2 : ab314490 IP in HAP1 (human chronic myelogenous leukemia near-haploid cell line) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314490 in HAP1 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-ACSS2 antibody [EPR27496-4] (<a href='/en-us/products/primary-antibodies/acss2-antibody-epr27496-4-ab314490'>ab314490</a>) at 1/30 dilution

All lanes:

HAP1 (human chronic myelogenous leukemia near-haploid cell line) 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: 8s

• WB

Supplier Data

Western blot - Anti-ACSS2 antibody [EPR27496-4] - BSA and Azide free (AB314491)

This data was developed using ab314490, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Performed under reducing conditions. In Western blot, ab314490 was shown to bind specifically to ACSS2. Target of interest was observed at 79 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in ACSS2 knockout cell line (lane 2). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-ACSS2 antibody [EPR27496-4] (<a href='/en-us/products/primary-antibodies/acss2-antibody-epr27496-4-ab314490'>ab314490</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell line) whole cell lysate at 50 µg

Lane 2:

ACSS2 knockout HAP1 whole cell lysate at 50 µg

Lane 3:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 50 µg

Lane 4:

L6 (rat skeletal muscle myoblast) whole cell lysate at 50 µg

Secondary

All lanes:

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

Observed band size: 79 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-ACSS2 antibody [EPR27496-4] - BSA and Azide free (AB314491)

This data was developed using ab314490, the same antibody clone in a different buffer formulation.

Western blot : Rabbit Monoclonal[EPR27496-4] to ACSS2 ab314490 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 75 kDa in Wild-type U-87 MG ab278079 cell lysates with no signal observed at this size in ACSS2 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-ACSS2 antibody [EPR27496-4] (<a href='/en-us/products/primary-antibodies/acss2-antibody-epr27496-4-ab314490'>ab314490</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG ab278079 at 20 µg

Lane 2:

ACSS2 knockout U-87 MG at 20 µg

Lane 3:

Wild-type HAP1 at 20 µg

Lane 4:

ACSS2 knockout HAP1 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 79 kDa

Observed band size: 75 kDa

false

• WB

Supplier Data

Western blot - Anti-ACSS2 antibody [EPR27496-4] - BSA and Azide free (AB314491)

This data was developed using ab314490, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : Lane 1 : 5 seconds, Lane 2-4 : 37 seconds.

All lanes:

Western blot - Anti-ACSS2 antibody [EPR27496-4] (<a href='/en-us/products/primary-antibodies/acss2-antibody-epr27496-4-ab314490'>ab314490</a>) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 50 µg

Lane 2:

Mouse brain tissue lysate at 50 µg

Lane 3:

Rat brain tissue lysate at 50 µg

Lane 4:

Rat liver tissue lysate at 50 µg

Secondary

All lanes:

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

Observed band size: 79 kDa

false