Anti-TOMM34 antibody [EPR30572-504]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOMM34 antibody [EPR30572-504] (AB324735)

Immunohistochemical analysis of paraffin-embedded Human small intestine tissue labeling TOMM34 with ab324735 at 1/50 (9.94 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : nearly no staining on human small intestine. The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOMM34 antibody [EPR30572-504] (AB324735)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling TOMM34 with ab324735 at 1/50 (9.94 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human testis. The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOMM34 antibody [EPR30572-504] (AB324735)

Immunohistochemical analysis of paraffin-embedded Clear cell carcinoma of human kidney tissue labeling TOMM34 with ab324735 at 1/50 (9.94 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on clear cell carcinoma of human kidney. The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-TOMM34 antibody [EPR30572-504] (AB324735)

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

Low expression : small intestine (PMID : 10101285).

Lanes 1-2 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross reactivity with human IgG at 1/2000 and lane 3 is applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000.

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

All lanes:

Western blot - Anti-TOMM34 antibody [EPR30572-504] (ab324735) at 1/2000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human small intestine tissue lysate at 20 µg

Lane 3:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

Lanes 1 - 2:

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

Lane 3:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 34 kDa,36 kDa

false

Exposure time: 125s

• WB

Supplier Data

Western blot - Anti-TOMM34 antibody [EPR30572-504] (AB324735)

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

Performed under reducing conditions.

In Western blot, ab324735 was shown to bind specifically to TOMM34. Target of interest was observed at 34 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in TOMM34 knockout cell line (lane 2) (lane 2, knockout cell line ab265546 / knockout cell lysate ab258730).

The identity of the higher MW band at approximately 50 kDa (in lane 2) is unknown.

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

Exposure time : Lanes 1-2 : 59 seconds; Lanes 3-4 : 26 seconds.

All lanes:

Western blot - Anti-TOMM34 antibody [EPR30572-504] (ab324735) at 1/2000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

TOMM34 knockout HeLa whole cell lysate at 20 µg

Lane 3:

293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 4:

293T transfected with siRNA specifically targeting TOMM34 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 34 kDa,36 kDa

false