Anti-Ten-1 antibody [EPR29567-35] - C-terminal

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ten-1 antibody [EPR29567-35] - C-terminal (AB325392)

Immunohistochemical analysis of paraffin-embedded Mouse cortex tissue labeling Ten-1(C-terminal) with ab325392 at 1/500 (1.05 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cortex. 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-Ten-1 antibody [EPR29567-35] - C-terminal (AB325392)

Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labeling Ten-1(C-terminal) with ab325392 at 1/500 (1.05 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse hippocampus (PMID : 12915301). 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-Ten-1 antibody [EPR29567-35] - C-terminal (AB325392)

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

Negative control : HeLa

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.Ten-1 can undergo degradation easily.

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

The band around 300 kDa is the full length form Ten-1 and 270 kDa the cleaved Ten-1(C-terminal)(PMID : 30618567).

All lanes:

Western blot - Anti-Ten-1 antibody [EPR29567-35] - C-terminal (ab325392) at 1/1000 dilution

Lane 1:

NCI-H69 (human lung small cell carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) 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 1/20000 dilution

Observed band size: 270-300 kDa,124 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-Ten-1 antibody [EPR29567-35] - C-terminal (AB325392)

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

Negative control : MCF7

Low expression : spleen

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Ten-1 antibody [EPR29567-35] - C-terminal (ab325392) at 1/1000 dilution

Lane 1:

A-673 (human muscle Ewings sarcoma cell line) whole cell lysate at 40 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 3:

Human cerebellum tissue lysate at 40 µg

Lane 4:

Human spleen tissue lysate at 40 µg

Secondary

All lanes:

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

Observed band size: 270-300 kDa,124 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Ten-1 antibody [EPR29567-35] - C-terminal (AB325392)

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

Low expression : spleen, lung

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

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

The band around 300 kDa is the full length form Ten-1 and 270 kDa the cleaved C-terminal Ten-1.

All lanes:

Western blot - Anti-Ten-1 antibody [EPR29567-35] - C-terminal (ab325392) at 1/1000 dilution

Lane 1:

Mouse hippocampus tissue lysate at 40 µg

Lane 2:

Mouse cerebellum tissue lysate at 40 µg

Lane 3:

Mouse spleen tissue lysate at 40 µg

Lane 4:

Mouse lung tissue lysate at 40 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 1/20000 dilution

Observed band size: 270-300 kDa,124 kDa

false

Exposure time: 15s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ten-1 antibody [EPR29567-35] - C-terminal (AB325392)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Ten-1(C-terminal) with ab325392 at 1/500 (1.05 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on mouse spleen. 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.

• I-ELISA

Supplier Data

Indirect ELISA - Anti-Ten-1 antibody [EPR29567-35] - C-terminal (AB325392)

Indirect ELISA analysis of ab325392 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : Human Ten-1 recombinant protein; Human Ten-2 recombinant protein; Human Ten-3 recombinant protein; Human Ten-4 recombinant protein.

Antigen concentration : 1000 ng/ml

This antibody does not cross-react with recombinant human Ten-2, Ten-3 or Ten-4 by Elisa.