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 (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

• 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

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

• 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.

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 with NFDM/TBST

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

• 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.

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 with NFDM/TBST

Lane 2:

TREM2 knockout THP-1 whole cell lysate at 20 µg with NFDM/TBST

Lane 3:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 4:

HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate at 20 µg with NFDM/TBST

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

• 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 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