Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (AB133672)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Methylmalonyl Coenzyme A mutase with Purified ab133672 at 1 : 100 dilution (9.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor®594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor®488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (AB133672)

Immunofluorescent staining of HeLa cells labelling Methylmalonyl Coenzyme A mutase with unpurified ab133672 at 1/100.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (AB133672)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling Methylmalonyl Coenzyme A mutase with Purified ab133672 at 1 : 100 dilution (8.7 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0)Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)was used as the secondary antibody.Negative control : PBS instead of the primary antibody.Hematoxylinwas used as a counterstain

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (AB133672)

Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling Methylmalonyl Coenzyme A mutase with unpurified ab133672 at 1/50.

• WB

Lab

Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (AB133672)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : Methylmalonyl Coenzyme A mutase knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lane 4 : K562 cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab133672 observed at 85 kDa. Red - loading control, ab8245, observed at 37 kDa.

Unpurified ab133672 was shown to specifically react with Methylmalonyl Coenzyme A mutase when Methylmalonyl Coenzyme A mutase knockout samples were used. Wild-type and Methylmalonyl Coenzyme A mutase knockout samples were subjected to SDS-PAGE. ab133672 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672)

Predicted band size: 83 kDa

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• WB

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Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (AB133672)

All lanes:

Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672) at 1/1000 dilution

Lane 1:

NIH 3T3 cell lysate at 10 µg

Lane 2:

K562 cell lysate at 10 µg

Lane 3:

293T cell lysate at 10 µg

Lane 4:

HeLa cell lysate at 10 µg

Lane 5:

Human fetal liver lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

Predicted band size: 83 kDa

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• WB

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Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (AB133672)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672) at 0.9 µg/mL

Lane 1:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 15 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 15 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 15 µg

Lane 4:

C6 (Rat glial tumor glial cell) whole cell lysates at 15 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: 83 kDa

false

• WB

Unknown

Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (AB133672)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Methylmalonyl Coenzyme A mutase antibody [EPR7739] (ab133672) at 0.9 µg/mL

Lane 1:

Mouse kidney lysates at 15 µg

Lane 2:

Rat kidney lysates at 15 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: 83 kDa

false