重组Anti-SCD1抗体[EPR21963] (ab236868)

All lanes : Anti-SCD1 antibody [EPR21963] (ab236868) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SCD knockout HeLa cell lysate
Lane 3 : HepG2 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 42 kDa

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.

 

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.

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).

All lanes : Anti-SCD1 antibody [EPR21963] (ab236868) at 1/1000 dilution

Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate
Lane 2 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate
Lane 3 : SK-MEL-28 (human malignant melanoma), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 42 kDa


Exposure time: 4 seconds

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.

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.

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. 

 

 

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.

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).

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).

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.

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.