Anti-LIGHT/TNFSF14 antibody [EPR29031-53]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LIGHT/TNFSF14 antibody [EPR29031-53] (AB323382)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling LIGHT/TNFSF14 with ab323382 at 1/200 (2.57 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Cytoplasmic staining on human spleen.
The section was incubated with ab323382 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LIGHT/TNFSF14 antibody [EPR29031-53] (AB323382)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling LIGHT/TNFSF14 with ab323382 at 1/200 (2.57 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Low expression : No staining on human skeletal muscle.
The section was incubated with ab323382 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LIGHT/TNFSF14 antibody [EPR29031-53] (AB323382)

Immunohistochemical analysis of paraffin-embedded Human endometrial carcinoma tissue labeling LIGHT/TNFSF14 with ab323382 at 1/200 (2.57 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Cytoplasmic staining on immunocytes of human endometrial carcinoma.
The section was incubated with ab323382 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LIGHT/TNFSF14 antibody [EPR29031-53] (AB323382)

Immunohistochemical analysis of paraffin-embedded Human lung adenocarcinoma tissue labeling LIGHT/TNFSF14 with ab323382 at 1/200 (2.57 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Cytoplasmic staining on on immunocytes of human lung adenocarcinoma.
The section was incubated with ab323382 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LIGHT/TNFSF14 antibody [EPR29031-53] (AB323382)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc, His-tag cell pellet. (B) HEK-293T transfected with a TNFSF14 expression vector containing a myc, His-tag cell pellet. tissue labeling LIGHT/TNFSF14 with ab323382 at 1/500 (1.028 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

No staining on (A) HEK-293T transfected with an empty vector containing a myc, His-tag cell pellet, positive staining on (B) HEK-293T transfected with a TNFSF14 expression vector containing a myc, His-tag cell pellet.
The section was incubated with ab323382 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LIGHT/TNFSF14 antibody [EPR29031-53] (AB323382)

Immunohistochemical analysis of paraffin-embedded (A) MCF7 (human breast adenocarcinoma epithelial cell) cell pellet. (B) No-GFP-CD16.NK-92 [CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag] cell pellet. tissue labeling LIGHT/TNFSF14 with ab323382 at 1/500 (1.028 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

No staining on (A) MCF7 cell pellet, positive staining on (B) No-GFP-CD16.NK-92 cell pellet.
The section was incubated with ab323382 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

Supplier Data

Western blot - Anti-LIGHT/TNFSF14 antibody [EPR29031-53] (AB323382)

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

Low expression : skeletal muscle.

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

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

All lanes:

Western blot - Anti-LIGHT/TNFSF14 antibody [EPR29031-53] (ab323382) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 18-28 kDa,36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-LIGHT/TNFSF14 antibody [EPR29031-53] (AB323382)

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

Low expression : MCF7, A549.

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

All lanes:

Western blot - Anti-LIGHT/TNFSF14 antibody [EPR29031-53] (ab323382) at 1/1000 dilution

Lane 1:

HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate at 80 µg

Lane 2:

No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) whole cell lysate at 80 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 80 µg

Lane 4:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 80 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: 18-28 kDa,124 kDa

false

Exposure time: 70s