Anti-SCD1 antibody [EPR21963]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SCD1 antibody [EPR21963] (AB236868)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling SCD1 with ab236868 at 1/100 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HepG2 cell line. Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution. Nuclear counterstain is DAPI.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SCD1 antibody [EPR21963] (AB236868)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell) cell line labeling SCD1 with ab236868 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SCD1 antibody [EPR21963] (AB236868)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-MEL-28 (Human malignant melanoma) cells labeling SCD1 with ab236868 at 1/100 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in SK-MEL-28 cell line. Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution. Nuclear counterstain is DAPI.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCD1 antibody [EPR21963] (AB236868)

Immunohistochemical analysis of paraffin-embedded human adipose tissue within cardiac muscle tissue labeling SCD1 with ab236868 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in human adipose cells in cardiac muscle (PMID : 15907797) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SCD1 antibody [EPR21963] (AB236868)

ab236868 staining SCD1 in wild-type HeLa cells, with negative expression in SCD1 knockout HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab236868 at 0.04 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 μg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

This product also work with 100% methanol (5 min) fixation under the same testing conditions.

• IP

Supplier Data

Immunoprecipitation - Anti-SCD1 antibody [EPR21963] (AB236868)

SCD1 was immunoprecipitated from 0.35 mg of HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate with ab236868 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab236868 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

Lane 1 : HEK-293 whole cell lysate 10 μg (input).
Lane 2 : ab236868 IP in HEK-293 whole cell lysate (+).
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab236868 in HEK-293 whole cell lysate (-).

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 30 seconds.

The expression profile observed is consistent with what has been described in the literature (PMID : 20876744; PMID : 9843580; PMID : 17449569). The full-length protein migrates at 37 kDa; the 28 kDa fragment may represent an SCD1 cleavage product.

All lanes:

Immunoprecipitation - Anti-SCD1 antibody [EPR21963] (ab236868)

Predicted band size: 42 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-SCD1 antibody [EPR21963] (AB236868)

SCD1 was immunoprecipitated from 10 μg of HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate with ab236868 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab236868 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

Lane 1 : HepG2 whole cell lysate 10 μg (input).
Lane 2 : ab236868 IP in HepG2 whole cell lysate (+).
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab236868 in HepG2 whole cell lysate (-).

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 30 seconds.

The expression profile observed is consistent with what has been described in the literature (PMID : 20876744; PMID : 9843580; PMID : 17449569). The full-length protein migrates at 37 kDa; the 28 kDa fragment may represent an SCD1 cleavage product.

All lanes:

Immunoprecipitation - Anti-SCD1 antibody [EPR21963] (ab236868)

Predicted band size: 42 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCD1 antibody [EPR21963] (AB236868)

Immunohistochemical analysis of paraffin-embedded mouse adipose tissue of stomach tissue labeling SCD1 with ab236868 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in adipose cells in mouse stomach (PMID : 11500518) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCD1 antibody [EPR21963] (AB236868)

Immunohistochemical analysis of paraffin-embedded rat adipose tissue of pancreas tissue labeling SCD1 with ab236868 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in adipose cells in rat pancreas (PMID : 11500518) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• WB

Lab

Western blot - Anti-SCD1 antibody [EPR21963] (AB236868)

anes 1-3 : Merged signal (red and green). Green - ab236868. Red - loading control ab8245 observed at 50 kDa.

ab236868 Anti-SCD1 antibody [EPR21963] was shown to specifically react with SCD1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265220 (knockout cell lysate ab257658) was used. Wild-type and SCD1 knockout samples were subjected to SDS-PAGE. ab236868 and Anti-tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-SCD1 antibody [EPR21963] (ab236868) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SCD knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SCD (SCD1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-scd-scd1-knockout-hela-cell-line-ab265220'>ab265220</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 42 kDa

false

• WB

Supplier Data

Western blot - Anti-SCD1 antibody [EPR21963] (AB236868)

Blocking/dilution buffer : 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID : 20876744; PMID : 9843580; PMID : 17449569). The full-length protein migrates at 37 kDa; the 28 kDa fragment may represent an SCD1 cleavage product.

We recommend that customers do not boil samples to prevent protein aggregation.

All lanes:

Western blot - Anti-SCD1 antibody [EPR21963] (ab236868) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

SK-MEL-28 (human malignant melanoma), 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 1/50000 dilution

Predicted band size: 42 kDa

false

Exposure time: 4s

• WB

Lab

Western blot - Anti-SCD1 antibody [EPR21963] (AB236868)

ab236868 was shown to react with SCD1 in wild-type HeLa cells in Western blot with loss of signal observed in SCD1 knockout cell line ab265220. Wild-type HeLa and SCD1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab236868 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/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-SCD1 antibody [EPR21963] (ab236868) at 1/1000 dilution

Lane 1:

Wild-type HeLa lysate at 35 µg

Lane 2:

SCD1 knock-out HeLa lysate at 35 µg

false