Anti-Sigma1-receptor antibody [EPR23266-69]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Sigma1-receptor antibody [EPR23266-69] (AB253192)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HT-29 (Human colorectal adenocarcinoma epithelial cell) cells labelling Sigma1-receptor with ab253192 at 1/600 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, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sigma1-receptor antibody [EPR23266-69] (AB253192)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Sigma1-receptor with ab253192 at 1/500 dilution (1.23 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human liver (PMID : 8954936). The section was incubated with ab253192 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sigma1-receptor antibody [EPR23266-69] (AB253192)

Immunohistochemical analysis of paraffin-embedded Human spinal cord tissue labeling Sigma1-receptor with ab253192 at 1/500 dilution (1.23 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human spinal cord (PMID : 23314020). The section was incubated with ab253192 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sigma1-receptor antibody [EPR23266-69] (AB253192)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Sigma1-receptor with ab253192 at 1/500 dilution (1.23 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat liver (PMID : 8954936). The section was incubated with ab253192 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sigma1-receptor antibody [EPR23266-69] (AB253192)

Immunohistochemical analysis of paraffin-embedded Rat spinal cord tissue labeling Sigma1-receptor with ab253192 at 1/500 dilution (1.23 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat spinal cord (PMID : 23314020). The section was incubated with ab253192 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• WB

Lab

Western blot - Anti-Sigma1-receptor antibody [EPR23266-69] (AB253192)

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

Exposure time : 3 minutes

All lanes:

Western blot - Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192) at 1/1000 dilution

Lane 1:

Caco-2 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

A375 (human malignant melanoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 25 kDa

Observed band size: 25 kDa

false

• WB

Collaborator

Western blot - Anti-Sigma1-receptor antibody [EPR23266-69] (AB253192)

ab253192 was shown to react with SIGMAR1 in wild-type HEK293T in Western blot with loss of signal observed in SIGMAR1 knockout cell line ab266619 (SIGMAR1 knockout cell lysate ab258666). Wild-type HEK293T and HeLa cell lysates and the corresponding SIGMAR1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab253192 overnight at 4 °C at a 1/5000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2μg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192) at 1/5000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human SIGMAR1 (Sigma1-receptor) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-sigmar1-sigma1-receptor-knockout-hek-293t-cell-lysate-ab258666'>ab258666</a>) at 20 µg

Lane 2:

Western blot - Human SIGMAR1 (Sigma1-receptor) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-sigmar1-sigma1-receptor-knockout-hek-293t-cell-line-ab266619'>ab266619</a>)

Lane 3:

Wild-type HeLa cell lysate at 20 µg

Lane 4:

SIGMAR1 knockout HeLa cell lysate at 20 µg

Secondary

All lanes:

goat anti-rabbit HRP at 0.2 µg/mL

Predicted band size: 25 kDa

false

• WB

Lab

Western blot - Anti-Sigma1-receptor antibody [EPR23266-69] (AB253192)

Lanes 1 - 4 : Merged signal (red and green). Green - ab253192 observed at 24 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab253192 was shown to react with Sigma1-receptor in wild-type HEK-293T cells in Western blot with loss of signal observed in SIGMAR1 knockout cell line ab266619 (SIGMAR1 knockout cell lysate ab258666). Wild-type HEK-293T and SIGMAR1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab253192 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human SIGMAR1 (Sigma1-receptor) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-sigmar1-sigma1-receptor-knockout-hek-293t-cell-lysate-ab258666'>ab258666</a>) at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Caco-2 cell lysate at 20 µg

Predicted band size: 25 kDa

Observed band size: 24 kDa,55 kDa

false

• WB

Lab

Western blot - Anti-Sigma1-receptor antibody [EPR23266-69] (AB253192)

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

Exposure time : 3 minutes

All lanes:

Western blot - Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat hippocampus tissue lysate at 20 µg

Lane 3:

Rat lung tissue lysate at 20 µg

Lane 4:

Rat kidney tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 25 kDa

Observed band size: 25 kDa

false

• WB

Lab

Western blot - Anti-Sigma1-receptor antibody [EPR23266-69] (AB253192)

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

Exposure time : 3.25 seconds

All lanes:

Western blot - Anti-Sigma1-receptor antibody [EPR23266-69] (ab253192) at 1/1000 dilution

Lane 1:

HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

HEK-293T transfected with Sigma1-receptor (WT) expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 25 kDa

Observed band size: 25 kDa

false