Anti-TM6SF2 antibody [EPR30567-58]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TM6SF2 antibody [EPR30567-58] (AB325553)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling TM6SF2 with ab325553 at 1/200 (2.47 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human liver. 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TM6SF2 antibody [EPR30567-58] (AB325553)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling TM6SF2 with ab325553 at 1/200 (2.47 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Low expression tissue : no staining on human testis (PMID : 24531328). 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TM6SF2 antibody [EPR30567-58] (AB325553)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a human TM6SF2 expression vector containing a Myc-His tag (B) HEK-293T transfected with empty vector containing a Myc-His tag tissue labeling TM6SF2 with ab325553 at 1/500 (0.988 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on HEK-293T transfected with a Myc-His-tagged human TM6SF2 construct, no staining on (B) HEK-293T transfected with empty vector containing a Myc-His tag. 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TM6SF2 antibody [EPR30567-58] (AB325553)

Immunohistochemical analysis of paraffin-embedded Human liver carcinoma tissue labeling TM6SF2 with ab325553 at 1/200 (2.47 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human liver carcinoma. 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TM6SF2 antibody [EPR30567-58] (AB325553)

Immunohistochemical analysis of paraffin-embedded (A) SK-OV-3 (human ovarian cancer epithelial cell) cell pellet (B) HEK-293T (human embryonic kidney epithelial cell) cell pellet tissue labeling TM6SF2 with ab325553 at 1/200 (2.47 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on SK-OV-3 (image A), no staining on HEK293T (image B). 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TM6SF2 antibody [EPR30567-58] (AB325553)

Immunohistochemical analysis of paraffin-embedded Human ileum tissue labeling TM6SF2 with ab325553 at 1/200 (2.47 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human ileum. 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• WB

Lab

Western blot - Anti-TM6SF2 antibody [EPR30567-58] (AB325553)

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

Low expression : HEK-293, testis(PMID : 24531328)

Samples are non-boiled as boiling may cause protein aggregation.

Lanes 1-4 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 and lanes 5-7 are incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

The identity of the band higher than 37 kDa is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lanes 1-4 : 158 seconds, lanes 5-7 : 37 seconds

All lanes:

Western blot - Anti-TM6SF2 antibody [EPR30567-58] (ab325553) at 1/1000 dilution

Lane 1:

SK-OV-3 (human ovarian cancer epithelial cell) transfected with scrambled siRNA control whole cell lysate at 40 µg

Lane 2:

SK-OV-3 transfected with siRNA specifically targeting TM6SF2 whole cell lysate at 40 µg

Lane 3:

SK-OV-3 whole cell lysate at 40 µg

Lane 4:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 40 µg

Lane 5:

Human small intestine tissue lysate at 40 µg

Lane 6:

Human testis tissue lysate at 40 µg

Lane 7:

Human liver cancer tissue lysate at 40 µg

Secondary

Lanes 1 - 4:

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 1/20000 dilution

Lanes 5 - 7:

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

Observed band size: 30-31 kDa,36 kDa

false