Anti-TMEM106B antibody [EPR30569-44] - C-terminal

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM106B antibody [EPR30569-44] - C-terminal (AB325607)

Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling TMEM106B with ab325607 at 1/500 (1.04 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human lung carcinoma.

The primary antibody was incubated 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-TMEM106B antibody [EPR30569-44] - C-terminal (AB325607)

Immunohistochemical analysis of paraffin-embedded Human Alzheimer's disease brain tissue labeling TMEM106B with ab325607 at 1/500 (1.04 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on Human Alzheimer's disease brain.

The primary antibody was incubated 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-TMEM106B antibody [EPR30569-44] - C-terminal (AB325607)

Immunohistochemical analysis of paraffin-embedded (A) PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) cell pellet and (B) Raji (human Burkitt's lymphoma B lymphocyte) cell pellet labeling TMEM106B with ab325607 at 1/1000 (0.52 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) PANC-1 cell pellet, no staining on (B) Raji cell pellet.

The primary antibody was incubated 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-TMEM106B antibody [EPR30569-44] - C-terminal (AB325607)

Immunohistochemical analysis of paraffin-embedded (A) Wild-type A549 (human lung carcinoma epithelial cell) cell pellet and (B) TMEM106B knockout A549 cell pellet labeling TMEM106B with ab325607 at 1/1000 (0.52 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) Wild-type A549 cell pellet, no staining on (B) TMEM106B knockout A549 cell pellet.

The primary antibody was incubated 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-TMEM106B antibody [EPR30569-44] - C-terminal (AB325607)

Immunohistochemical analysis of paraffin-embedded Human endometrial carcinoma tissue labeling TMEM106B with ab325607 at 1/500 (1.04 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human endometrial carcinoma.

The primary antibody was incubated 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-TMEM106B antibody [EPR30569-44] - C-terminal (AB325607)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling TMEM106B with ab325607 at 1/500 (1.04 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human cerebrum (PMID : 24252750).

The primary antibody was incubated 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

Lab

Multiplex immunohistochemistry - Anti-TMEM106B antibody [EPR30569-44] - C-terminal (AB325607)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human cerebrum tissue staining TMEM106B with ab325607 at a 1 : 500 (1.04 µg/ml) dilution, ab108597 anti-LAMP1 used at 1 : 400 (0.37 µg/ml) dilution and ab300140 anti-P2Y12 used at a 1 : 40000 (0.013 µg/ml) dilution.

Panel A : merged staining of anti-TMEM106B (green; Opal™520), anti-LAMP1 (magenta; Opal™570) and anti-P2Y12 (yellow; Opal™690) on human cerebrum.
Panel B : anti-TMEM106B showed granular staining in human cerebrum.
Panel C : anti-LAMP1 staining lysosome in human cerebrum.
Panel D : anti-P2Y12 staining microglia 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 ab325607, ab108597 and ab300140 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM106B antibody [EPR30569-44] - C-terminal (AB325607)

Immunohistochemical analysis of paraffin-embedded Mouse Alzheimer's disease brain tissue labeling TMEM106B with ab325607 at 1/2000 (0.26 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on Mouse Alzheimer's disease brain.

The primary antibody was incubated 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-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-TMEM106B antibody [EPR30569-44] - C-terminal (AB325607)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (perfused fixed) tissue labeling TMEM106B with ab325607 at 1/50 (10.4 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 µg/mL dilution (Green).

Panel A : merged staining of anti-TMEM106B (ab325607, green), anti-NeuN (ab190565, magenta) on mouse cerebrum. Panel B : anti-TMEM106B stained on mouse cerebrum. Panel C : anti-NeuN stained in neurons of mouse cerebrum. The section was incubated in two rounds of staining : in the order of ab325607 and ab190565, for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMEM106B antibody [EPR30569-44] - C-terminal (AB325607)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling TMEM106B with ab325607 at 1/2000 (0.26 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum.

The primary antibody was incubated 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-TMEM106B antibody [EPR30569-44] - C-terminal (AB325607)

Immunohistochemical analysis of paraffin-embedded Mouse lung carcinoma tissue labeling TMEM106B with ab325607 at 1/2000 (0.26 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung carcinoma.

The primary antibody was incubated 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

• WB

Lab

Western blot - Anti-TMEM106B antibody [EPR30569-44] - C-terminal (AB325607)

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

This antibody does not cross-react with human TMEM106C and human TMEM106A by western blot.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa);Anti-Myc tag® antibody (1 : 5000) (40KDa);.

All lanes:

Western blot - Anti-TMEM106B antibody [EPR30569-44] - C-terminal (ab325607) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 2:

293T transfected with a human TMEM106B expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T transfected with a human TMEM106C expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 4:

293T transfected with a human TMEM106A expression vector containing a myc-His-tag® 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: 40 kDa,36 kDa

false

Exposure time: 8s

• WB

Lab

Western blot - Anti-TMEM106B antibody [EPR30569-44] - C-terminal (AB325607)

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

Negative control : Raji

Performed under reducing conditions.

In Western blot, ab325607 was shown to bind specifically to TMEM106B. Target of interest was observed at 40 and 30 kDa in wild-type A549 cell lysates (lane 1) with no signal observed at this size in TMEM106B knockout cell line (lane 2, knockout cell line ab273711 / knockout cell lysate ab273769).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 39709600).

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

The identity of the bands higher than 150 kDa is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-TMEM106B antibody [EPR30569-44] - C-terminal (ab325607) at 1/1000 dilution

Lane 1:

Wild-type A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

TMEM106B knockout A549 whole cell lysate at 20 µg

Lane 3:

AsPC-1 (human Pancreas epithelial cell) whole cell lysate at 20 µg

Lane 4:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 6:

3T3-L1 (mouse embryonic fibroblast) 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,40 kDa,36 kDa

true

Exposure time: 103s