ET1611-64

CPT2 Recombinant Rabbit Monoclonal Antibody [SN06-70]

cat.: ET1611-64

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	SN06-70

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 74 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human CPT2 aa 609-658 / 658.
Positive control:	Hela cell lysate, 293 cell lysate, HepG2 cell lysate, NIH/3T3 cell lysate, SK-Br-3, SW480, human liver carcinoma tissue, human kidney tissue, mouse kidney tissue, human muscle tissue, human colon carcinoma tissue, HeLa.
Subcellular location:	Mitochondrion inner membrane, nucleolus, nucleoplasm.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:1,000 1:50-1:200 1:50-1:200 1:1,000 1:1,000
Uniprot #:	SwissProt: P23786 Human \| P52825 Mouse \| P18886 Rat
Alternative names:	Carnitine O palmitoyltransferase 2 Carnitine O palmitoyltransferase 2 mitochondrial Carnitine O-palmitoyltransferase 2 Carnitine palmitoyltransferase 2 Carnitine palmitoyltransferase II CPT 1 CPT 2 CPT II CPT1 CPT2 CPT2_HUMAN CPTASE CPTII IIAE4 mitochondrial

Images

Fig1: Western blot analysis of CPT2 on different lysates with Rabbit anti-CPT2 antibody (ET1611-64) at 1/1,000 dilution.

Lane 1: PC-12 cell lysate
Lane 2: Mouse kidney tissue lysate
Lane 3: Rat kidney tissue lysate

Cell lysates/proteins at 20 µg/Lane.
Tisssue lysates/proteins at 40 µg/Lane.

Predicted band size: 74 kDa
Observed band size: 67 kDa

Exposure time: 25 seconds;

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 (ET1611-64) at 1/1,000 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 CPT2 on different lysates with Rabbit anti-CPT2 antibody (ET1611-64) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-CPT2 KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 74 kDa
Observed band size: 67 kDa

Exposure time: 60 seconds; 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 (ET1611-64) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: Western blot analysis of CPT2 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1611-64, 1/1,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: Hela cell lysate Lane 2: 293 cell lysate Lane 3: HepG2 cell lysate Lane 4: NIH/3T3 cell lysate
	Fig4: Immunocytochemistry analysis of SK-Br-3 cells labeling CPT2 with Rabbit anti-CPT2 antibody (ET1611-64) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-CPT2 antibody (ET1611-64) at 1/50 dilution in 2% negative goat serum 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. Nuclear DNA was labelled in blue with DAPI.
	Fig5: ICC staining of CPT2 in SW480 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-64, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

	Fig6: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-CPT2 antibody (ET1611-64) at 1/1,000 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 (ET1611-64) at 1/1,000 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 mouse kidney tissue with Rabbit anti-CPT2 antibody (ET1611-64) at 1/1,000 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 (ET1611-64) at 1/1,000 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 human muscle tissue with Rabbit anti-CPT2 antibody (ET1611-64) at 1/1,000 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 (ET1611-64) at 1/1,000 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 human colon carcinoma tissue with Rabbit anti-CPT2 antibody (ET1611-64) at 1/1,000 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 (ET1611-64) at 1/1,000 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.

Fig10: Flow cytometric analysis of HeLa cells labeling CPT2.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1611-64, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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