Anti-MTHFR antibody [EPR19781]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MTHFR antibody [EPR19781] (AB203786)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling MTHFR with ab203786 at 1/100 dilution followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining on Jurkat cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab203786 at 1/100 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTHFR antibody [EPR19781] (AB203786)

Immunohistochemical analysis of paraffin-embedded Human thyroid carcinoma tissue labeling MTHFR with ab203786 at 1/1000 (0.509 µg/mL) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human thyroid carcinoma. The section was incubated with ab203786 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution) for 20 minutes.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTHFR antibody [EPR19781] (AB203786)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling MTHFR with ab203786 at 1/1000 (0.509 µg/mL) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human kidney. The section was incubated with ab203786 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution) for 20 minutes.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MTHFR antibody [EPR19781] (AB203786)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling MTHFR with ab203786 at 1/100 dilution followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining on RAW 264.7 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab203786 at 1/100 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-MTHFR antibody [EPR19781] (AB203786)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 : 3 minutes; Lane 2,3 and 4 : 30 seconds.

All lanes:

Western blot - Anti-MTHFR antibody [EPR19781] (ab203786) at 1/1000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Lane 3:

Human fetal kidney lysate at 10 µg

Lane 4:

Human spleen lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 35 kDa,75 kDa

Observed band size: 75 kDa

false

• WB

Supplier Data

Western blot - Anti-MTHFR antibody [EPR19781] (AB203786)

Blocking/Dilution buffer : 5% NFDM/TBST.

Lanes 1 - 3:

Western blot - Anti-MTHFR antibody [EPR19781] (ab203786) at 1/1000 dilution

Lanes 4 - 5:

Western blot - Anti-MTHFR antibody [EPR19781] (ab203786) at 1/5000 dilution

Lane 1:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 2:

HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 3:

Human fetal liver tissue lysate at 20 µg

Lane 4:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 5:

Mouse placenta 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 1/100000 dilution

Predicted band size: 75 kDa

Observed band size: 75 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-MTHFR antibody [EPR19781] (AB203786)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-MTHFR antibody [EPR19781] (ab203786) at 1/1000 dilution

Lane 1:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 20 µg

Lane 3:

Mouse 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-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 42 kDa,45 kDa,75 kDa

Observed band size: 42 kDa,75 kDa,90-120 kDa

false

Exposure time: 3min

• IP

Supplier Data

Immunoprecipitation - Anti-MTHFR antibody [EPR19781] (AB203786)

MTHFR was immunoprecipitated from 0.35mg of HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate with ab203786 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab203786 at 1/500 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : HT-29 whole cell lysate, 10μg (Input).

Lane 2 : ab203786 IP in HT-29 whole cell lysate.

Lane 3 : Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab203786 in HT-29 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-MTHFR antibody [EPR19781] (ab203786)

Predicted band size: 75 kDa

false