Anti-Sigma1-receptor antibody [EPR25145-11] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sigma1-receptor antibody [EPR25145-11] - BSA and Azide free (AB307549)

This data was developed using ab307548, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Sigma1-receptor with ab307548 at 1/1000 (0.533 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human liver (PMID : 29399701).The section was incubated with ab307548 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) was used. 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-Sigma1-receptor antibody [EPR25145-11] - BSA and Azide free (AB307549)

This data was developed using ab307548, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded (A) Wild-type HEK-29 tissue labeling Sigma1-receptor with ab307548 at 1/1000 (0.533 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on (A) Wild-type HEK-293T cell pellet, no staining on (B) SIGMAR1 knockout HEK-293T (ab266619) cell pellet.The section was incubated with ab307548 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) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Sigma1-receptor antibody [EPR25145-11] - BSA and Azide free (AB307549)

This data was developed using ab307548, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Wild-type 293T (human embryonic kidney epithelial cell, Right) / Sigma1-receptor knockout 293T (Left) cells labelling Sigma1-receptor with ab307548 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Positive staining on 293T cells (ab255449), while no staining on Sigma1-receptor knockout 293T cells (ab266619).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Sigma1-receptor antibody [EPR25145-11] - BSA and Azide free (AB307549)

This data was developed using ab307548, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C6 (rat glial tumor glial cell) cells labelling Sigma1-receptor with ab307548 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sigma1-receptor antibody [EPR25145-11] - BSA and Azide free (AB307549)

This data was developed using ab307548, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Sigma1-receptor with ab307548 at 1/1000 (0.533 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse cerebrum (PMID : 31596232).The section was incubated with ab307548 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) was used. 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-Sigma1-receptor antibody [EPR25145-11] - BSA and Azide free (AB307549)

This data was developed using ab307548, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Sigma1-receptor with ab307548 at 1/1000 (0.533 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on rat liver.The section was incubated with ab307548 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) was used. 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-Sigma1-receptor antibody [EPR25145-11] - BSA and Azide free (AB307549)

This data was developed using ab307548, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Sigma1-receptor with ab307548 at 1/1000 (0.533 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse liver. The section was incubated with ab307548 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) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Sigma1-receptor antibody [EPR25145-11] - BSA and Azide free (AB307549)

This data was developed using ab307548, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Sigma1-receptor with ab307548 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• WB

Supplier Data

Western blot - Anti-Sigma1-receptor antibody [EPR25145-11] - BSA and Azide free (AB307549)

This data was developed using 307548, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : Low expression : brain (PMID : 28350844), spleen (PMID : 28350844). Exposure time :

All lanes:

Western blot - Anti-Sigma1-receptor antibody [EPR25145-11] (<a href='/en-us/products/primary-antibodies/sigma1-receptor-antibody-epr25145-11-ab307548'>ab307548</a>) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate 20 μg

Lane 2:

Mouse brain tissue lysate 20 μg

Lane 3:

Rat liver tissue lysate 20 μg

Lane 4:

Rat brain tissue lysate 20 μg

Lane 5:

Human liver tissue lysate 20 μg

Lane 6:

Human spleen tissue lysate 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: 26 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-Sigma1-receptor antibody [EPR25145-11] - BSA and Azide free (AB307549)

This data was developed using 307548, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Performed under reducing conditions. In Western blot, ab307548 was shown to bind specifically to SIGMAR1. A band was observed at 26 kDa in wild-type 293T cell lysates with whereas no signal observed at this size in SIGMAR1 knockout cell line ab266619 (knockout cell lysate ab258666). Exposure time : Lanes 1-2 : 180 seconds; Lane 3 : 136 seconds.

All lanes:

Western blot - Anti-Sigma1-receptor antibody [EPR25145-11] (<a href='/en-us/products/primary-antibodies/sigma1-receptor-antibody-epr25145-11-ab307548'>ab307548</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

SIGMAR1 knockout HEK293T whole cell lysate at 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 6:

C6 (rat glial tumor glial 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 1/100000 dilution

Observed band size: 26 kDa

false

Exposure time: 180s