Anti-ACAT1 antibody [EPR28035-8] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACAT1 antibody [EPR28035-8] - BSA and Azide free (AB312321)

This data was developed using ab312320, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling ACAT1 with ab312320 at 1/5000 (0.103 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon. The section was incubated with ab312320 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). 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-ACAT1 antibody [EPR28035-8] - BSA and Azide free (AB312321)

This data was developed using ab312320, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human thyroid carcin tissue labeling ACAT1 with ab312320 at 1/5000 (0.103 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human thyroid carcinoma. The section was incubated with ab312320 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). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ACAT1 antibody [EPR28035-8] - BSA and Azide free (AB312321)

This data was developed using ab312320, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A549 (human lung carcinoma epithelial cell) cells labelling ACAT1 with ab312320 at 1/500 (1.034 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing mitochondrial staining in A549 cell line.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/1000 1ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ACAT1 antibody [EPR28035-8] - BSA and Azide free (AB312321)

This data was developed using ab312320, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A549 (human lung carcinoma epithelial cell) cells labelling ACAT1 with ab312320 at 1/500 dilution (0.1 ug)/Right (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 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACAT1 antibody [EPR28035-8] - BSA and Azide free (AB312321)

This data was developed using ab312320, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse squamous cell tissue labeling ACAT1 with ab312320 at 1/5000 (0.103 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse squamous cell carcinoma. The section was incubated with ab312320 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). 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-ACAT1 antibody [EPR28035-8] - BSA and Azide free (AB312321)

This data was developed using ab312320, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling ACAT1 with ab312320 at 1/5000 (0.103 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat kidney. The section was incubated with ab312320 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). 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-ACAT1 antibody [EPR28035-8] - BSA and Azide free (AB312321)

This data was developed using ab312320, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling ACAT1 with ab312320 at 1/5000 (0.103 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse kidney. The section was incubated with ab312320 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). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ACAT1 antibody [EPR28035-8] - BSA and Azide free (AB312321)

This data was developed using ab312320, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling ACAT1 with ab312320 at 1/500 (1.034 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing mitochondrial staining in NIH/3T3 cell line.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/1000 1ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-ACAT1 antibody [EPR28035-8] - BSA and Azide free (AB312321)

This data was developed using ab312320, 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 ACAT1 with ab312320 at 1/500 dilution (0.1 ug)/Red (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 1/5000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-ACAT1 antibody [EPR28035-8] - BSA and Azide free (AB312321)

This data was developed using ab312320, the same antibody clone in a different buffer formulation. ACAT1 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab312320 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab312320 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate Lane 2 : ab312320 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab312320 in NIH/3T3 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 10 seconds

All lanes:

Immunoprecipitation - Anti-ACAT1 antibody [EPR28035-8] (<a href='/en-us/products/primary-antibodies/acat1-antibody-epr28035-8-ab312320'>ab312320</a>) at 1/30 dilution

All lanes:

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

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-ACAT1 antibody [EPR28035-8] - BSA and Azide free (AB312321)

This data was developed using ab312320, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Anti-TOMM20 antibody [EPR15581-54] (ab186735). In Western blot, anti-TOMM20 antibody [EPR15581-54] (ab186735) staining at 1/1000 dilution. Anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution. Exposure time : 10 seconds.

All lanes:

Western blot - Anti-ACAT1 antibody [EPR28035-8] (<a href='/en-us/products/primary-antibodies/acat1-antibody-epr28035-8-ab312320'>ab312320</a>) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) non-mitochondrial fraction at 20 µg

Lane 2:

NIH/3T3 mitochondria fraction at 20 µg

Lane 3:

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

Lane 4:

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

Lane 5:

A549 (human lu carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

PC-3 (human prostate adenocarcinoma epithelial 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: 45 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-ACAT1 antibody [EPR28035-8] - BSA and Azide free (AB312321)

This data was developed using ab312320, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution. Exposure time : 8 seconds.

All lanes:

Western blot - Anti-ACAT1 antibody [EPR28035-8] (<a href='/en-us/products/primary-antibodies/acat1-antibody-epr28035-8-ab312320'>ab312320</a>) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

NIH/3T3 transfected with siRNA specifically targeti ACAT1 whole cell lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Rat liver tissue 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: 45 kDa

false

Exposure time: 8s