Anti-TREM2 antibody [EPR26209-22]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TREM2 antibody [EPR26209-22] (AB318262)

Immunohistochemical analysis of paraffin-embedded Human Alzheimer's cerebrum tissue labeling TREM2 with ab318262 at 1/100 (5.29 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human Alzheimer's cerebrum (PMID : 25186950; : 25186950; PMID : 28592261 ). The section was incubated with ab318262 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-TREM2 antibody [EPR26209-22] (AB318262)

Immunohistochemical analysis of paraffin-embedded Human glioblastoma tissue labeling TREM2 with ab318262 at 1/100 (5.29 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human glioblastoma. The section was incubated with ab318262 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

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-TREM2 antibody [EPR26209-22] (AB318262)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human cerebrum tissue staining TREM2 with ab318262 at a 1 : 100 (5.29 ug/ml) dilution, ab306583 anti-TMEM119 used at 1 : 2000 (0.255 ug/ml) dilution and ab218309 anti-GFAP used at a 1 : 1000 (1.325 ug/ml) dilution.

Panel A : merged staining of anti-TREM2 (green; Opal™520), anti-TMEM119 (magenta; Opal™690) and anti-GFAP (yellow; Opal™570) on human cerebrum.
Panel B : anti-TREM2 staining microglia in human cerebrum.
Panel C : anti-TMEM119 staining microglia in human cerebrum.
Panel D : anti-GFAP staining astrocytes in human cerebrum.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab318262, ab306583 and ab218309 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-TREM2 antibody [EPR26209-22] (AB318262)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human astrocytoma tissue staining TREM2 with ab318262 at a 1 : 100 (5.29 ug/ml) dilution, ab306583 anti-TMEM119 used at 1 : 2000 (0.255 ug/ml) dilution and ab218309 anti-GFAP used at a 1 : 1000 (1.325 ug/ml) dilution.

Panel A : merged staining of anti-TREM2 (green; Opal™520), anti-TMEM119 (magenta; Opal™690) and anti-GFAP (yellow; Opal™570) on human astrocytoma.
Panel B : anti-TREM2 staining microglia in human astrocytoma.
Panel C : anti-TMEM119 staining microglia in human astrocytoma.
Panel D : anti-GFAP staining astrocyte in human astrocytoma.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab318262, ab306583 and ab218309 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval 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-TREM2 antibody [EPR26209-22] (AB318262)

Immunohistochemical analysis of paraffin-embedded Human astrocytoma tissue labeling TREM2 with ab318262 at 1/100 (5.29 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human astrocytoma. The section was incubated with ab318262 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TREM2 antibody [EPR26209-22] (AB318262)

Immunohistochemical analysis of formalin fixed paraffin embedded human glioblastoma labelling TREM2 with ab318262 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins ab318262 anti-TREM2 antibody [EPR26209-22] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TREM2 antibody [EPR26209-22] (AB318262)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling TREM2 with ab318262 at 1/100 (5.29 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human liver (PMID : 12472885). The section was incubated with ab318262 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-TREM2 antibody [EPR26209-22] (AB318262)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a TREM2 expression vector containing a Myc-His tag. (B) HEK-293T transfected with TREM1 expression vector containing a His tag. (C) HEK-293T transfected with empty vector containing a Myc-His tag. tissue labeling TREM2 with ab318262 at 1/2000 (0.265 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a TREM2 expression vector containing a Myc-His tag, negative staining on (B) HEK-293T transfected with TREM1 expression vector containing a His tag and (C) HEK-293T transfected with empty vector containing a Myc-His tag. The section was incubated with ab318262 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-TREM2 antibody [EPR26209-22] (AB318262)

Immunohistochemical analysis of paraffin-embedded (A) THP-1 (human monocytic leukemia monocyte) cell pellet (B) SH-SY5Y (human neuroblastoma epithelial cell) cell pellet (C) HL-60 (human acute promyelocytic leukemia promyeloblast) cell pellet tissue labeling TREM2 with ab318262 at 1/100 (5.29 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) THP-1 cell pellet, negative staining on (B) SH-SY5Y cell pellet and (C) HL-60 cell pellet. The section was incubated with ab318262 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

Supplier Data

Immunoprecipitation - Anti-TREM2 antibody [EPR26209-22] (AB318262)

TREM2 was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte) whole cell lysate with ab318262 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab318262 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

Lane 1 : THP-1 (human monocytic leukemia monocyte) whole cell lysate
Lane 2 : ab318262 IP in THP-1 (human monocytic leukemia monocyte) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab318262 in THP-1 whole cell lysate

All lanes:

Immunoprecipitation - Anti-TREM2 antibody [EPR26209-22] (ab318262) 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='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 84s

• IP

Supplier Data

Immunoprecipitation - Anti-TREM2 antibody [EPR26209-22] (AB318262)

Immunoprecipitation of TREM2 in THP-1 cells. Lysates were prepared and immunoprecipitation was performed using 2μg of ab318262 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Immunoprecipitation - Anti-TREM2 antibody [EPR26209-22] (ab318262) at 2 µg

All lanes:

THP-1 cell lysates

Observed band size: 31 kDa

false

• WB

Supplier Data

Western blot - Anti-TREM2 antibody [EPR26209-22] (AB318262)

ab318262 was shown to react with TREM2 in wild-type THP-1 cells in Western blot with loss of signal observed in TREM2 knockout cell line ab269489. 10 μg of concentrated culture media from PMA-treated Wild-type THP-1 and TREM2 knockout cell lysates. Wild-type THP-1 and TREM2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab318262 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-TREM2 antibody [EPR26209-22] (ab318262) at 1/1000 dilution

Lane 1:

10 μg of concentrated culture media from PMA-treated Wild-type THP-1 lysate at 20 µg

Lane 2:

10 μg of concentrated culture media from PMA-treated TREM2 knock-out THP-1 lysate at 20 µg

Lane 2:

Western blot - Human TREM2 knockout THP-1 cell line (<a href='/en-us/products/cell-lines/human-trem2-knockout-thp-1-cell-line-ab269489'>ab269489</a>)

Observed band size: 31 kDa

false

• WB

Supplier Data

Western blot - Anti-TREM2 antibody [EPR26209-22] (AB318262)

In Western blot, ab318262 was shown to bind specifically to TREM2. Target of interest was observed at 30 kDa in wild-type THP-1 cell lysates (lane 1) with no signal observed at this size in TREM2 knockout cell line (lane 2) (lane 2, knockout cell line ab269489 / knockout cell lysate ab269652).

Negative control : SH-SY5Y, HL-60.

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

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

All lanes:

Western blot - Anti-TREM2 antibody [EPR26209-22] (ab318262) at 1/1000 dilution

Lane 1:

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

Lane 2:

TREM2 knockout THP-1 whole cell lysate at 20 µg

Lane 3:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HL-60 (human acute promyelocytic leukemia promyeloblast) 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: 30 kDa,36 kDa

false

Exposure time: 180s