Anti-MYSM1 antibody [EPR18657] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-MYSM1 antibody [EPR18657] - BSA and Azide free (AB251165)

This data was developed using ab193081, the same antibody clone in a different buffer formulation.

MYSM1 was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab193081 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab193081 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : HeLa whole cell lysate 10μg (Input).

Lane 2 : ab193081 IP in HeLa whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab193081 in HeLa whole cell lysate.

Predicted band size : 66kDa, 95kDa.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

Two bands detected represent two isoforms.

All lanes:

Immunoprecipitation - Anti-MYSM1 antibody [EPR18657] (<a href='/en-us/products/primary-antibodies/mysm1-antibody-epr18657-ab193081'>ab193081</a>)

Predicted band size: 95 kDa,66 kDa

true

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-MYSM1 antibody [EPR18657] - BSA and Azide free (AB251165)

This data was developed using ab193081, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Two bands detected represent two isoforms.

All lanes:

Western blot - Anti-MYSM1 antibody [EPR18657] (<a href='/en-us/products/primary-antibodies/mysm1-antibody-epr18657-ab193081'>ab193081</a>) at 1/1000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 3:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 4:

NCCIT (Human pluripotent embryonic carcinoma cell line) whole cell lysate at 20 µg

Lane 5:

K562 (Human chronic myelogenous leukemia cell line from bone marrow) 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

Predicted band size: 95 kDa

Observed band size: 66 kDa,95 kDa

true

Exposure time: 1min

• WB

Lab

Western blot - Anti-MYSM1 antibody [EPR18657] - BSA and Azide free (AB251165)

This data was developed using ab193081, the same antibody clone in a different buffer formulation.

False colour image of Western blot : Anti-MYSM1 antibody [EPR18657] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab193081 was shown to bind specifically to MYSM1. A band was observed at 110 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in MYSM1 knockout cell line ab267297 (knockout cell lysate ab257548). To generate this image, wild-type and MYSM1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-MYSM1 antibody [EPR18657] (<a href='/en-us/products/primary-antibodies/mysm1-antibody-epr18657-ab193081'>ab193081</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

MYSM1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human MYSM1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-mysm1-knockout-hek-293t-cell-line-ab267297'>ab267297</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

RT-4 cell lysate at 20 µg

Predicted band size: 95 kDa

Observed band size: 110 kDa

false

• WB

Supplier Data

Western blot - Anti-MYSM1 antibody [EPR18657] - BSA and Azide free (AB251165)

This data was developed using ab193081, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-MYSM1 antibody [EPR18657] (<a href='/en-us/products/primary-antibodies/mysm1-antibody-epr18657-ab193081'>ab193081</a>) at 1/1000 dilution

Lane 1:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 2:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast cell line) 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

Predicted band size: 95 kDa

Observed band size: 95 kDa

true

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-MYSM1 antibody [EPR18657] - BSA and Azide free (AB251165)

This data was developed using ab193081, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-MYSM1 antibody [EPR18657] (<a href='/en-us/products/primary-antibodies/mysm1-antibody-epr18657-ab193081'>ab193081</a>) at 1/1000 dilution

All lanes:

mESC (Mouse embryonic stem cell line) 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

Predicted band size: 95 kDa

Observed band size: 95 kDa

true

Exposure time: 8s