Anti-MTHFD1 antibody [EPR29810-508]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTHFD1 antibody [EPR29810-508] (AB324379)

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling MTHFD1 with ab324379 at 1/1000 (0.49 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon carcinoma.
The primary antibody was incubated for 30 mins 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTHFD1 antibody [EPR29810-508] (AB324379)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling MTHFD1 with ab324379 at 1/1000 (0.49 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human pancreas.
The primary antibody was incubated for 30 mins 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTHFD1 antibody [EPR29810-508] (AB324379)

Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma and adjacent liver tissue labeling MTHFD1 with ab324379 at 1/1000 (0.49 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining observed in adjacent normal human liver tissue, with reduced expression of MTHFD1 in hepatocellular carcinoma tissue (PMID : 30997850).
The primary antibody was incubated for 30 mins 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MTHFD1 antibody [EPR29810-508] (AB324379)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293 (human embryonic kidney epithelial cell) cells labelling MTHFD1 with ab324379 at 1/100 (4.9 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ab150081 1000 2ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in HEK-293 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/ab7291 1000 1ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab324379 at 1/100 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution. -ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTHFD1 antibody [EPR29810-508] (AB324379)

Immunohistochemical analysis of paraffin-embedded (A) Normal mouse liver. (B) Mouse hepatocellular carcinoma. tissue labeling MTHFD1 with ab324379 at 1/1000 (0.49 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining observed in (A) normal mouse liver, with reduced expression of MTHFD1 in (B) hepatocellular carcinoma tissue.
The primary antibody was incubated for 30 mins 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-MTHFD1 antibody [EPR29810-508] (AB324379)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-MTHFD1 antibody [EPR29810-508] (ab324379) at 1/1000 dilution

Lane 1:

HEK-293 (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HEK-293 transfected with siRNA specifically targeting MTHFD1 whole cell 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

Observed band size: 101 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-MTHFD1 antibody [EPR29810-508] (AB324379)

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

Lanes 1-5 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 and lane 6 is incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

The identity of the band at approximately 60 kDa in lane 3 is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-2, 5-6 : 15 seconds; Lanes 3-4 : 70 seconds

All lanes:

Western blot - Anti-MTHFD1 antibody [EPR29810-508] (ab324379) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 6:

Human liver tissue lysate at 20 µg

Secondary

Lanes 1 - 5:

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 1/2000 dilution

Lane 6:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

Observed band size: 101 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-MTHFD1 antibody [EPR29810-508] (AB324379)

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

This antibody does not cross-react with human MTHFD1L by western blot.

In Western blot, Anti-6X His tag® antibody [AD1.1.10] - (ab15149) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-MTHFD1 antibody [EPR29810-508] (ab324379) at 1/1000 dilution

Lane 1:

No-tagged human MTHFD1 fragment at 10 ng

Lane 2:

His-tagged human MTHFD1L fragment at 10 ng

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

Observed band size: 50 kDa,30 kDa

false

Exposure time: 15s