Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free

• IP

Unknown

Immunoprecipitation - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (AB274378)

HDAC8 was immunoprecipitated from 0.35 mg K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab274372 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab274372 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate 10 ug.

Lane 2 : ab274372 IP in K-562 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab274372 in K-562 whole cell lysate

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

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274372).

All lanes:

Immunoprecipitation - Anti-HDAC8 antibody [EPR23837-45] (<a href='/en-us/products/primary-antibodies/hdac8-antibody-epr23837-45-ab274372'>ab274372</a>)

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (AB274378)

HDAC8 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab274372 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab274372 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug.

Lane 2 : ab274372 IP in NIH/3T3 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab274372 in NIH/3T3 whole cell lysate.

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

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274372).

All lanes:

Immunoprecipitation - Anti-HDAC8 antibody [EPR23837-45] (<a href='/en-us/products/primary-antibodies/hdac8-antibody-epr23837-45-ab274372'>ab274372</a>)

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

• WB

Lab

Western blot - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (AB274378)

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

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274372)

All lanes:

Western blot - Anti-HDAC8 antibody [EPR23837-45] (<a href='/en-us/products/primary-antibodies/hdac8-antibody-epr23837-45-ab274372'>ab274372</a>) at 1/1000 dilution

Lane 1:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

LADMAC (mouse bone marrow monocyte macrophage) whole cell lysate at 20 µg

Lane 4:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

• WB

Lab

Western blot - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (AB274378)

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

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 3 minutes

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274372)

All lanes:

Western blot - Anti-HDAC8 antibody [EPR23837-45] (<a href='/en-us/products/primary-antibodies/hdac8-antibody-epr23837-45-ab274372'>ab274372</a>) at 1/1000 dilution

All lanes:

Human brain tissue lysate at 20 µg

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

• WB

Lab

Western blot - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (AB274378)

This data was developed using ab274372, the same antibody clone in a different buffer formulation.

Western blot : Rabbit Monoclonal [EPR23837-45] to HDAC8 ab274372 staining at 1/1000 dilution, shown in green; Mouse anti-CANX (ab238078) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 42 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in HDAC8 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 5pc 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-HDAC8 antibody [EPR23837-45] (<a href='/en-us/products/primary-antibodies/hdac8-antibody-epr23837-45-ab274372'>ab274372</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG at 20 µg

Lane 2:

HDAC8 knockout U-87 MG at 20 µg

Lane 3:

HeLa at 20 µg

Lane 4:

HCT 116 at 20 µg

Secondary

All lanes:

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

Predicted band size: 42 kDa

Observed band size: 42 kDa

false