Anti-S1P1/EDG1 antibody [EPR21202]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-S1P1/EDG1 antibody [EPR21202] (AB233386)

ab233386 was shown to react with S1PR1 in wild-type SKHEP1 cells in immunocytochemistry with loss of signal observed in a S1PR1 knockout cell line. Wild-type and knockout cells were mixed and pelleted at a 1 : 1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with ab233386 at 1/400dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 µg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).

This data was 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S1P1/EDG1 antibody [EPR21202] (AB233386)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling S1P1/EDG1 with ab233386 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining in endothelial cells of human colon (PMID : 25588843) is observed. Counterstained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S1P1/EDG1 antibody [EPR21202] (AB233386)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling S1P1/EDG1 with ab233386 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining in endothelial cells of human kidney (PMID : 25588843) is observed. Counterstained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• WB

Lab

Western blot - Anti-S1P1/EDG1 antibody [EPR21202] (AB233386)

Blocking buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-S1P1/EDG1 antibody [EPR21202] (ab233386) at 1/1000 dilution

Lanes 1 and 3:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lanes 2 and 4:

BxPC-3 (human pancreas adenocarcinoma cell line) 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

Predicted band size: 43 kDa

false

Exposure time: 114s

• WB

Lab

Western blot - Anti-S1P1/EDG1 antibody [EPR21202] (AB233386)

ab233386 was shown to react with S1PR1 in wild-type SKHEP1 cells in Western blot with loss of signal observed in a S1PR1 knockout cell line. Wild-type SKHEP1 and S1PR1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab233386 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2ug/mL before imaging.

This data was 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-S1P1/EDG1 antibody [EPR21202] (ab233386) at 1/1000 dilution

Lane 1:

Wild-type SKHEP1 lysate at 20 µg

Lane 2:

S1PR1 knock-out SKHEP1 lysate at 20 µg

Observed band size: 43 kDa

false