Anti-TIPE2 抗体 [EPR27964-2]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-TIPE2 antibody [EPR27964-2] (AB316848)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labelling TIPE2 with ab316848 at 1/50 (10.04 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 cytoplasmic and nuclear staining in THP-1 cell line.
Negative control : HepG2 (PMID : 24274578).
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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIPE2 antibody [EPR27964-2] (AB316848)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling TIPE2 with ab316848 at 1/100 (5.02 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil.
The section was incubated with ab316848 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIPE2 antibody [EPR27964-2] (AB316848)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling TIPE2 with ab316848 at 1/100 (5.02 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on stromal cells of human colon.
The section was incubated with ab316848 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIPE2 antibody [EPR27964-2] (AB316848)

Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling TIPE2 with ab316848 at 1/100 (5.02 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human lung carcinoma (PMID : 25946186).
The section was incubated with ab316848 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-TIPE2 antibody [EPR27964-2] (AB316848)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling TIPE2 with ab316848 at 1/100 (5.02 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : Nearly no staining on human testis.
The section was incubated with ab316848 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

• IP

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Immunoprecipitation - Anti-TIPE2 antibody [EPR27964-2] (AB316848)

TIPE2 was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte) whole cell lysate with ab316848 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316848 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : THP-1 (human monocytic leukemia monocyte) whole cell lysate
Lane 2 : ab316848 IP in THP-1 (human monocytic leukemia monocyte) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316848 in THP-1 whole cell lysate

All lanes:

Immunoprecipitation - Anti-TIPE2 antibody [EPR27964-2] (ab316848) at 1/30 dilution

All lanes:

THP-1 (human monocytic leukemia monocyte) whole cell lysate

Secondary

All lanes:

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

false

Exposure time: 180s

• WB

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Western blot - Anti-TIPE2 antibody [EPR27964-2] (AB316848)

Negative control : HepG2 (PMID : 24274578), 293T (PMID : 27696294).

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

All lanes:

Western blot - Anti-TIPE2 antibody [EPR27964-2] (ab316848) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

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

Secondary

All lanes:

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

Observed band size: 18 kDa,36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-TIPE2 antibody [EPR27964-2] (AB316848)

Low expression : testis (PMID : 30638932).

The identity of the bands between 25 kDa and 75 kDa are unknown.

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

All lanes:

Western blot - Anti-TIPE2 antibody [EPR27964-2] (ab316848) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 20 µg

Lane 2:

Human testis tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 18 kDa

true

Exposure time: 103s