Anti-CPT1A antibody [8F6AE9]

• Supplier: Abcam
• Catalog: ab128568
• Prices: $390.00
• Sizes: 100 µg
• Host: Mouse
• Clonality: Monoclonal
• Isotype: IgG2b
• Clone: 8F6AE9
• Applications: IHC-P WB ICC/IF ICC/IF ELISA
• Species Reactivities: Mouse, Rat, Human
• Antigen: Recombinant fragment, corresponding to amino acids 489-773 of Human CPT1A
• Description: Mouse Monoclonal
• Gene: CPT1A
• Conjugate: Unconjugated

Product images

All lanes : Anti-CPT1A antibody [8F6AE9] (ab128568) at 1 µg/mlLane 1 : MarkerLane 2 : Partial Human Recombinant CPT1A protein (ab128569) at 0.1 µgLane 3 : Full Length Human Recombinant OTC protein at 0.1 µgLane 4 : HepG2 (Human hepatocellular carcinoma cell line) Whole Cell Lysate at 20 µgLane 5 : H9C2 (Rat cardiomyoblast cell line) Whole Cell Lysate at 20 µgLane 6 : H4IIE ( Rat hepatoma cell line) Whole Cell Lysate at 20 µgLane 7 : RHM (Rat heart mitochondrial homogenate) at 20 µgLane 8 : HLH (Human liver homogenate) at 20 µgLane 9 : RLH (Rat liver homogenate) at 20 µgLane 10 : MLH (Mouse liver homogenate) at 20 µgSecondaryGoat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

Immunofluorescent staining of CPT1A in H9C2 cells (rat) using ab128568.Fixation: 4% paraformaldehyde PBS fixed for 20 minutes Permebilization: 0.1% Triton X-100 PBS for 30 minutes at room temperature while rockingBlocking: 2x Sigma Block 0.1% Triton X-100 H2O for 2 hours at room temperature while rocking Primary antibodies: Anti-CPT1A antibody (ab128568) 0.5 ug/mL 1x Sigma Block with 0.1% Triton X-100 incubated overnight at 4 °C.Washing: 3x 1% NGS 10 minutes/wash. Secondary antibodies: Alexa 488 GAM 1:1000 diluted in 1% NGS with 0.1% Triton X-100 PBS incubated for 2 hours at room temperature while rocking.Washing: 3x 1% NGS 10 minutes/wash. DAPI: 20 ng/mL in 1% NGS, 0.1% Triton X-100 PBS.Washing: 1x 1% NGS 10 minutes/wash.

Overlay histogram showing HeLa cells stained with ab128568 (red line) or no primary antibody (black line). The cells were fixed with 4% paraformaldehyde (15 min) and then permeabilized with 0.1% Triton X-100 in PBS, 3% BSA for 10 min. The cells were then incubated in 3% BSA in PBS for 10 minutes to block non-specific protein-protein interactions followed by the antibody (ab128568, 1µg/mL) for 60 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96871) at 1/1000 dilution for 30 min.

All lanes : Anti-CPT1A antibody [8F6AE9] (ab128568) at 1 µg/mlLane 1 : MarkerLane 2 : HHH (Human heart homogenate) RIPA Extract Immunoprecipitated with no primary antibodyLane 3 : HHH (Human heart homogenate) RIPA Extract Immunoprecipitated with Anti-CPT1A antibody 8F6AE9 (ab128568)Lane 4 : HHH (Human heart homogenate) RIPA Extract 20 µgSecondaryGoat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1:10000developed using the ECL techniquePerformed under non-reducing conditions.

IHC image of CPT1A staining in human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab128568, 0.5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

In-Cell ELISA for Anti-CPT1A antibody (ab128568) stained HeLa cells (human)Seeding: HeLa cells seeded in a 1:2 dilution series starting at 60,000 cells/well across Row A, 30,000 cells/well across Row B, etc. Row H contains no cells. Table: Example In-Cell ELISA Average Data from shown plate.Fixation: 4% paraformaldehyde PBS fixed for 15 minutes Permeabilization: 0.3% Triton X-100 PBS for 30 minutes at room temperature while shakingBlocking: 2x Sigma Block 0.3% Triton X-100 H2O for 2 hours at room temperature while shaking Primary antibodies: All primaries diluted in 1x Sigma Block with 0.3% Triton X-100 incubated overnight at 4 °C. • Columns 1-3: Anti-CPT1A antibody (ab128568) 10 ug/mL • Columns 4-6: Anti-CPT1A antibody (ab128568) 1 ug/mL • Columns 7-9: Anti-CPT1A antibody (ab128568) 0.1 ug/mL • Columns 10-12: No PrimaryWashing: Briefly 4x with 0.3% TWEEN-20 PBS Secondary antibodies: Licor GAM 800 1:1000 in 1x Sigma Block 0.3% Triton X-100 H2O incubated for 2 hours at room temperature

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