Anti-MTR antibody

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-MTR antibody (AB66039)

ICC/IF image of ab66039 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66039, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa, Hek293 and HepG2 cells at 5µg/ml, and in 4% PFA fixed (10 min) MCF7 cells at 5µg/ml.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTR antibody (AB66039)

IHC image of ab66039 staining in pancreas formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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 ab66039, 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.

• WB

Project5584****

Western blot - Anti-MTR antibody (AB66039)

All lanes:

Western blot - Anti-MTR antibody (ab66039) at 1 µg/mL

Lane 1:

TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate at 10 µg

Lane 2:

Liver (Mouse) Tissue Lysate at 10 µg

Lane 3:

Thymus (Rat) Tissue Lysate at 10 µg

Lane 4:

Pancreas (Mouse) Tissue Lysate at 10 µg

Lane 5:

Jurkat Whole Cell Lysate - Staurosporine Treated (24hr, 500nM) at 10 µg

Lane 6:

HeLa Whole Cell Lysate - Staurosporine Treated (24hr, 500nM) at 10 µg

Lane 7:

HeLa Whole Cell Lysate - Bleomycin Treated (20U/ml) at 10 µg

Lane 8:

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

Lane 9:

Kidney (Mouse) Tissue Lysate at 10 µg

Lane 10:

Liver (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 140 kDa

Observed band size: 117 kDa,141 kDa,35 kDa,50 kDa,55 kDa,65 kDa,85 kDa

false

• WB

CiteAb

Western blot - Anti-MTR antibody (AB66039)

MTR western blot using anti-MTR antibody ab66039. Publication image and figure legend from Pérez-Sepúlveda, A., España-Perrot, P. P., et al., 2013, Biomed Res Int, PubMed 24024209.

ab66039 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab66039 please see the product overview.

(a) MTHFR and (b) MTR protein levels analyzed from placental tissue by western blot. Values are ± SEM from 32 controls and 16 PE patients.

false