Anti-mtTFA antibody - Mitochondrial Marker

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody - Mitochondrial Marker (AB47517)

ab47517 staining mtTFA in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab47517 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• WB

Ap25807****

Western blot - Anti-mtTFA antibody - Mitochondrial Marker (AB47517)

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab47517 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-mtTFA antibody - Mitochondrial Marker (ab47517) at 1 µg/mL

Lane 1:

Skeletal Muscle (Human) Tissue Lysate - adult normal tissue at 20 µg

Lane 2:

Human heart tissue lysate - total protein (<a href='/en-us/products/unavailable/human-heart-tissue-lysate-total-protein-ab29431'>ab29431</a>) at 20 µg

Lane 3:

Brown adipose (Mouse) Tissue Lysate at 20 µg

Lane 4:

Brown adipose (Rat) Tissue Lysate at 20 µg

Lane 5:

Heart (Mouse) Tissue Lysate at 20 µg

Lane 6:

Heart (Rat) Tissue Lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

Predicted band size: 29 kDa

Observed band size: 100 kDa,18 kDa,29 kDa

true

Exposure time: 8min

• WB

Project5922****

Western blot - Anti-mtTFA antibody - Mitochondrial Marker (AB47517)

All lanes:

Western blot - Anti-mtTFA antibody - Mitochondrial Marker (ab47517) at 2 µg/mL

Lane 1:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 20 µg

Lane 2:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 20 µg with Human mtTFA peptide (<a href='/en-us/products/unavailable/human-mttfa-peptide-ab47516'>ab47516</a>)

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 29 kDa

Observed band size: 24 kDa,26 kDa,50 kDa

false

Exposure time: 10min

• WB

Project

Western blot - Anti-mtTFA antibody - Mitochondrial Marker (AB47517)

All lanes:

Western blot - Anti-mtTFA antibody - Mitochondrial Marker (ab47517) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Lane 3:

A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 29 kDa

Observed band size: 25 kDa,60 kDa

false