Anti-TREM2 antibody [EPR26210-1]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TREM2 antibody [EPR26210-1] (AB305103)

Immunohistochemical analysis of paraffin-embedded (A) RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cell pellet and (B) Neuro-2a (mouse neuroblastoma neuroblast) cell pellet, labeling TREM2 with ab305103 at 1/500 dilution (1.014 µg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection) kit.

The section was incubated with ab305103 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.

Positive staining in RAW264.7 cell pellet was observed with no staining in Neuro-2a cell pellet. Secondary antibody only control : RAW264.7 cell pellet with only ready to use Leica DS9800 (Bond™ Polymer Refine Detection) kit.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TREM2 antibody [EPR26210-1] (AB305103)

Immunohistochemical analysis of paraffin-embedded tissue labeling Trem2 with ab305103 at 1/500 dilution, followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining on (A) whole brain tissue up to postnatal day 5 (P5) from wild-type C57BL/6JGpt mice and no staining on (B) whole brain tissue up to postnatal day 5 (P5) from TREM2 knockout mice
The section was incubated with ab305103 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.
The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Trem2-KO homozygous mice (Strain ID : T003346).

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

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 [EPR26210-1] (AB305103)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T cells transfected with a mouse TREM2 expression vector containing a GFP tag and (B) HEK-293T cells transfected with an empty vector containing a GFP tag (labeling TREM2 with ab305103 at 1/500 (1.014 µg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on (A) HEK-293T cells transfected with a Trem2 expression vector containing a GFP tag. No staining on (B) HEK-293T cells transfected with empty vector containing a GFP tag. The section was incubated with ab305103 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 Leica DS9800 (Bond™ Polymer Refine Detection) kit. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TREM2 antibody [EPR26210-1] (AB305103)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) cells labelling TREM2 with ab305103 at 1/500 (1.014 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 staining on RAW 264.7 cell line.Negative control : Neuro-2a (PMID : 19302484).Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TREM2 antibody [EPR26210-1] (AB305103)

Immunohistochemical analysis of paraffin-embedded (A) FAD4T Transgenic mouse Alzheimer’s disease cerebrum and (B) Mouse normal cerebrum (labeling TREM2 with ab305103 at 1/500 (1.014 µg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on (A) Mouse Alzheimer’s disease cerebrum, no staining on (B) Mouse normal cerebrum. The section was incubated with ab305103 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 Leica DS9800 (Bond™ Polymer Refine Detection) kit. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

• WB

Supplier Data

Western blot - Anti-TREM2 antibody [EPR26210-1] (AB305103)

Blocking and diluting buffer and concentration : 5% NFDM/TBST TREM2 is a glycoprotein of approximately 35-50 kDa and detected as a 25 kDa band after treated with Protein Deglycosylation MIX II. The identity of the higher MW band at approximately 70 kDa is unknown.

All lanes:

Western blot - Anti-TREM2 antibody [EPR26210-1] (ab305103) at 1/1000 dilution

Lane 1:

Untreated J774A.1 (mouse reticum cell sarcoma monocyte macrophage), whole cell lysate at 30 µg

Lane 2:

J774A.1 whole cell lysate treated with Protein Deglycosylation Mix II at 30 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

Predicted band size: 35-50 kDa,25 kDa

Observed band size: 35-50 kDa,25 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-TREM2 antibody [EPR26210-1] (AB305103)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression profile and molecular weight observed are consistent with what has been described in the literature (PMID : 29611543; PMID : 28559417). Negative control : L929, F9 (PMID : 11241283), Neuro-2a (PMID : 19302484). The identity of the higher MW band at approximately 70 kDa is unknown.

All lanes:

Western blot - Anti-TREM2 antibody [EPR26210-1] (ab305103) at 1/1000 dilution

Lane 1:

J774A.1 (mouse reticum cell sarcoma monocyte macrophage), whole cell lysate at 20 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

Lane 3:

N9 (mouse microglia), whole cell lysate at 20 µg

Lane 4:

L929 (mouse connective tissue fibroblast), whole cell lysate at 20 µg

Lane 5:

Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate at 20 µg

Lane 6:

F9 (mouse embryonal carcinoma epithelial cell), 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

Predicted band size: 25 kDa

Observed band size: 35-50 kDa

false

Exposure time: 3min