Anti-CPT2 antibody [EPR13626] - C-terminal

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT2 antibody [EPR13626] - C-terminal (AB181114)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling CPT2 with unpurified ab181114 at 1/100 dilution, followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CPT2 antibody [EPR13626] - C-terminal (AB181114)

Immunofluorescent analysis of 4% paraformaldehyde-fixed MCF7 cells labeling CPT2 with unpurified ab181114 at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody at 1/200 dilution. Counter stained with Dapi.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT2 antibody [EPR13626] - C-terminal (AB181114)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling CPT2 with unpurified ab181114 at 1/100 dilution, followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT2 antibody [EPR13626] - C-terminal (AB181114)

Immunohistochemical analysis of paraffin embedded human liver carcinoma tissue section labelling CPT2 with purified ab181114 at dilution of 1/50. The secondary antibody used was Goat Anti-Rabbit IgG H&L (HRP) (ab97051), at a dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CPT2 antibody [EPR13626] - C-terminal (AB181114)

Immunocytochemistry/Immunofluorescence analysis of MCF-7 (human breast carcinoma) cells labelling CPT2 with purified ab181114 at 1/100. Cells were fixed with 100% methanol and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000) using ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary. Nuclei couterstained with DAPI (blue).

For negative control 1, rabbit primary antibody was used, followed by anti-mouse secondary antibody (ab150120). For negative control 2, mouse primary antibody (ab7291) was used followed by anti-rabbit secondary antibody (ab150077).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT2 antibody [EPR13626] - C-terminal (AB181114)

Immunohistochemical analysis of paraffin embedded rat colon tissue section labelling CPT2 with purified ab181114 at dilution of 1/50. The secondary antibody used was HRP-conjugated Goat Anti-Rabbit IgG H&L (ab97051) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPT2 antibody [EPR13626] - C-terminal (AB181114)

Immunohistochemical analysis of paraffin embedded mouse kidney tissue section labelling CPT2 with purified ab181114 at dilution of 1/50. The secondary antibody used was HRP-conjugated Goat Anti-Rabbit IgG H&L (ab97051) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

• WB

Lab

Western blot - Anti-CPT2 antibody [EPR13626] - C-terminal (AB181114)

Lanes 1 - 4 : Merged signal (red and green). Green - ab181114 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab181114 was shown to specifically react with CPT2 in wild-type HAP1 cells. No band was observed when CPT2 knockout samples were examined. Wild-type and CPT2 knockout samples were subjected to SDS-PAGE. ab181114 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CPT2 antibody [EPR13626] - C-terminal (ab181114) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

CPT2 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Lane 4:

MCF7 whole cell lysate at 20 µg

Predicted band size: 74 kDa

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• WB

Lab

Western blot - Anti-CPT2 antibody [EPR13626] - C-terminal (AB181114)

Lanes 1- 2 : Merged signal (red and green). Green - ab181114 observed at 74 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab181114 was shown to react with CPT2/CPT1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265931 (knockout cell lysate ab257180) was used. Wild-type HeLa and CPT2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab181114 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CPT2 antibody [EPR13626] - C-terminal (ab181114) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CPT2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CPT2 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cpt2-knockout-hela-cell-line-ab265931'>ab265931</a>)

Predicted band size: 74 kDa

Observed band size: 74 kDa

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• WB

Unknown

Western blot - Anti-CPT2 antibody [EPR13626] - C-terminal (AB181114)

Blocking and diluting buffer 5% NFDM/TBST

All lanes:

Western blot - Anti-CPT2 antibody [EPR13626] - C-terminal (ab181114) at 1/2000 dilution

Lane 1:

Human fetal kidney tissue lysate at 20 µg

Lane 2:

MCF-7 (human breast carcinoma) whole cell lysate at 20 µg

Lane 3:

Mouse heart tissue lysate at 20 µg

Lane 4:

Mouse kidney tissue lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 74 kDa

Observed band size: 67 kDa

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• WB

Unknown

Western blot - Anti-CPT2 antibody [EPR13626] - C-terminal (AB181114)

Blocking and diluting buffer 5% NFDM/TBST

All lanes:

Western blot - Anti-CPT2 antibody [EPR13626] - C-terminal (ab181114) at 1/1000 dilution

Lane 1:

Rat kidney tissue lysate at 20 µg

Lane 2:

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 1/20000 dilution

Predicted band size: 74 kDa

Observed band size: 67 kDa

false

• WB

Supplier Data

Western blot - Anti-CPT2 antibody [EPR13626] - C-terminal (AB181114)

All lanes:

Western blot - Anti-CPT2 antibody [EPR13626] - C-terminal (ab181114) at 1/10000 dilution

Lane 1:

Human fetal liver tissue lysate at 20 µg

Lane 2:

Human fetal kidney 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-ab136636'>ab136636</a>) at 1/500 dilution

Predicted band size: 74 kDa

Observed band size: 67 kDa

false