Anti-MTAP antibody [EPR22574-104]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR22574-104] (AB252934)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling MTAP with ab252934 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in stroma cells and no staining in tumor cells of human breast cancer (PMID : 26751376) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR22574-104] (AB252934)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling MTAP with ab252934 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in human breast cancer (PMID : 26751376,18712977) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR22574-104] (AB252934)

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling MTAP with ab252934 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in stroma cells and no staining in tumor cells of human bladder cancer (PMID : 27270441) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR22574-104] (AB252934)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling MTAP with ab252934 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in human breast (PMID : 26751376, 18712977) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IP

Unknown

Immunoprecipitation - Anti-MTAP antibody [EPR22574-104] (AB252934)

MTAP was immunoprecipitated from 0.35 mg of HT-29 (human colorectal adenocarcinoma cell line) whole cell lysate with ab252934 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab252934 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

Lane 1 : HT-29 whole cell lysate 10 μg (Input).
Lane 2 : ab252934 IP in HT-29 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab252934 in HT-29 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-MTAP antibody [EPR22574-104] (ab252934)

Predicted band size: 31 kDa

Observed band size: 32 kDa

false

• WB

Unknown

Western blot - Anti-MTAP antibody [EPR22574-104] (AB252934)

Blocking and dilution buffer : 5% NFDM/TBST.

Negative control : A549 (PMID : 17548352).

The expression molecular weight observed is consistent with what has been described in the literature (PMID : 15492751).

All lanes:

Western blot - Anti-MTAP antibody [EPR22574-104] (ab252934) at 1/1000 dilution

Lane 1:

HT-29 (human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

MDA-MB-235S (human ductal carcinoma cell line) whole cell lysate at 20 µg

Lane 3:

A549 (human lung carcinoma 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: 31 kDa

Observed band size: 32 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-MTAP antibody [EPR22574-104] (AB252934)

Western blot : Anti-MTAP antibody [EPR22574-104] ab252934 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 31kDa in Wild-type HeLa cell lysates with no signal observed at this size in MTAP knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% 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 and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-MTAP antibody [EPR22574-104] (ab252934) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysates at 20 µg

Lane 2:

MTAP knockout HeLa cell lysates at 20 µg

Lane 3:

HepG2 cell lysates at 20 µg

Lane 4:

A549 cell lysates at 20 µg

Secondary

All lanes:

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

Predicted band size: 31 kDa

Observed band size: 31 kDa

false

• WB

Unknown

Western blot - Anti-MTAP antibody [EPR22574-104] (AB252934)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-MTAP antibody [EPR22574-104] (ab252934) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell line) whole cell lysate at 10 µg

Lane 2:

NIH/3T3 (mouse embryo 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: 31 kDa

Observed band size: 32 kDa

false

Exposure time: 3min