Anti-Tissue Factor antibody [RM1198]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tissue Factor antibody [RM1198] (AB318995)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Tissue Factor with ab318995 at 1/2000 (0.249 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human kidney. The section was incubated with ab318995 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-Tissue Factor antibody [RM1198] (AB318995)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling Tissue Factor with ab318995 at 1/2000 (0.249 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human skeletal muscle. The section was incubated with ab318995 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-Tissue Factor antibody [RM1198] (AB318995)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Tween-20 permeabilized A431 (human epidermoid carcinoma epithelial cell) cells labelling Tissue Factor with ab318995 at 1/100 (4.97 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous and cytoplasmic staining in A431 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : MCF7 (PMID : 36980251).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Tissue Factor antibody [RM1198] (AB318995)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A431 (human epidermoid carcinoma epithelial cell) (Right) / MCF7 (human breast adenocarcinoma epithelial cell) (Left) cells labelling Tissue Factor with ab318995 at 1/5000 dilution (0.1ug) / Magenta compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : MCF7.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tissue Factor antibody [RM1198] (AB318995)

Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labeling Tissue Factor with ab318995 at 1/2000 (0.249 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human cervical carcinoma. The section was incubated with ab318995 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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Tissue Factor antibody [RM1198] (AB318995)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Tissue Factor with ab318995 at 1/50 dilution (1ug) / Magenta compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Tissue Factor antibody [RM1198] (AB318995)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Tween-20 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Tissue Factor with ab318995 at 1/100 (4.97 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous and cytoplasmic staining in NIH/3T3 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).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IP

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Immunoprecipitation - Anti-Tissue Factor antibody [RM1198] (AB318995)

Tissue Factor was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab318995 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab318995 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 2 : ab318995 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab318995 in NIH/3T3 whole cell lysate

All lanes:

Immunoprecipitation - Anti-Tissue Factor antibody [RM1198] (ab318995) at 1/30 dilution

All lanes:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/ko/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 24s

• WB

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Western blot - Anti-Tissue Factor antibody [RM1198] (AB318995)

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-Tissue Factor antibody [RM1198] (ab318995) at 1/1000 dilution

Lane 1:

Wide-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 60 µg

Lane 2:

Tissue Factor knockout HAP1 whole cell lysate at 60 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 47 kDa,36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Tissue Factor antibody [RM1198] (AB318995)

Low expression : skeletal muscle.

Lane 1 and 2 were developed using a high sensitivity ECL substrate.

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

Exposure time : Lane 1-2 : 180 seconds; Lane 3-4 : 114 seconds

All lanes:

Western blot - Anti-Tissue Factor antibody [RM1198] (ab318995) at 1/1000 dilution

Lane 1:

Human hippocampus tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 47 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Tissue Factor antibody [RM1198] (AB318995)

Low expression : MCF-7 (PMID : 24137414, PMID : 28938620).

Lane 4 was developed using a high sensitivity ECL substrate.

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

Exposure time : Lane 1 : 180 seconds; Lane 2 : 26 seconds; Lane 3 : 148 seconds; Lane 4 : 70 seconds

All lanes:

Western blot - Anti-Tissue Factor antibody [RM1198] (ab318995) at 1/1000 dilution

Lane 1:

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

Lane 2:

BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 47 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Tissue Factor antibody [RM1198] (AB318995)

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

All lanes:

Western blot - Anti-Tissue Factor antibody [RM1198] (ab318995) at 1/1000 dilution

Lane 1:

B16-F10 (mouse skin melanoma cell) whole cell lysate at 20 µg

Lane 2:

A20 (mouse reticulum sarcoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 47 kDa,36 kDa

false

Exposure time: 180s