HA721093

ACAT-1 Recombinant Rabbit Monoclonal Antibody [JE64-40]

cat.: HA721093

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell
Clonality:	Monoclonal
Clone number:	JE64-40

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 45 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within human ACAT1 aa 378-427/427.
Positive control:	Raji cell lysate, HepG2 cell lysate, rat liver tissue lysate, rat heart tissue lysate, HepG2, human liver tissue, human liver carcinoma tissue, rat liver tissue, mouse liver tissue, mouse heart tissue.
Subcellular location:	Mitochondrion.
Recommended Dilutions: WB IHC-P IF-Cell	1:500-1:1,000 1:100-1:400 1:250
Uniprot #:	SwissProt: P24752 Human \| Q8QZT1 Mouse \| P17764 Rat
Alternative names:	ACAT 1 ACAT acat1 Acetoacetyl CoA thiolase acetoacetyl Coenzyme A thiolase Acetoacetyl-CoA thiolase Acetyl CoA acetyltransferase, mitochondrial Acetyl Coenzyme A acetyltransferase 1 Acetyl-CoA acetyltransferase acetyl-coa acetyltransferase precursor, mitochondrial Acetyl-CoA thiolase, mitochondrial acetyl-Coenzyme A acetyltransferase 1 MAT mitochondrial acetoacetyl-CoA thiolase mitochondrial RATACAL T2 testicular tissue protein Li 198 THIL THIL_HUMAN

Images

Fig1: Western blot analysis of ACAT-1 on different lysates with Rabbit anti-ACAT-1 antibody (HA721093) at 1/2,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si ACAT-1 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 45 kDa
Observed band size: 42 kDa

Exposure time: 1 minute; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721093) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of ACAT-1 on different lysates with Rabbit anti-ACAT-1 antibody (HA721093) at 1/500 dilution.

Lane 1: Raji cell lysate
Lane 2: HepG2 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 45 kDa
Observed band size: 42 kDa

Exposure time: 4 minutes;

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721093) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

	Fig3: Western blot analysis of ACAT-1 on different lysates with Rabbit anti-ACAT-1 antibody (HA721093) at 1/500 dilution. Lane 1: Rat liver tissue lysate Lane 2: Rat heart tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 45 kDa Observed band size: 42 kDa Exposure time: 1 minutes; 8% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721093) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
	Fig4: Immunocytochemistry analysis of HepG2 cells labeling ACAT-1 with Rabbit anti-ACAT-1 antibody (HA721093) at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ACAT-1 antibody (HA721093) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.
	Fig5: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-ACAT-1 antibody (HA721093) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721093) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig6: Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue with Rabbit anti-ACAT-1 antibody (HA721093) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721093) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig7: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-ACAT-1 antibody (HA721093) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721093) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig8: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-ACAT-1 antibody (HA721093) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721093) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig9: Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-ACAT-1 antibody (HA721093) at 1/400 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721093) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.