Anti-Pentraxin 3/PTX3 antibody [EPR30744-540]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pentraxin 3/PTX3 antibody [EPR30744-540] (AB325639)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a human PTX3 expression vector containing a Myc-His tag and (B) HEK-293T transfected with empty vector containing a Myc-His tag tissue labeling Pentraxin 3/PTX3 with ab325639 at 1/2000 (0.254 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a human PTX3 expression vector containing a Myc-His tag, no staining on (B) HEK-293T transfected with empty vector containing a Myc-His tag. 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-Pentraxin 3/PTX3 antibody [EPR30744-540] (AB325639)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling Pentraxin 3/PTX3 with ab325639 at 1/100 (5.07 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on stroma of mouse skeletal muscle (PMID : 27900389). 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-Pentraxin 3/PTX3 antibody [EPR30744-540] (AB325639)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling Pentraxin 3/PTX3 with ab325639 at 1/100 (5.07 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on stroma of mouse cardiac muscle (PMID : 31209269; PMID : 23372656). 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-Pentraxin 3/PTX3 antibody [EPR30744-540] (AB325639)

Immunohistochemical analysis of paraffin-embedded (A) 3T3-L1 (mouse embryonic fibroblast) cell pellet (B) NIH/3T3 (mouse embryonic fibroblast) cell pellet (C) B16-F10 (mouse skin melanoma cell) cell pellet (D) Neuro-2a (mouse neuroblastoma cell) cell pellet labeling Pentraxin 3/PTX3 with ab325639 at 1/100 (5.07 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) 3T3-L1 cell pellet, (B) NIH/3T3 cell pellet, no staining on (C) B16-F10 cell pellet, (D) Neuro-2a 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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Pentraxin 3/PTX3 antibody [EPR30744-540] (AB325639)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse lung treated with lps (1ug/ml 16h)+bfa (1ug/ml 16h) (+) ex vivo tissue staining Pentraxin 3/PTX3 with ab325639 at a 1 : 100 (5.07 µg/ml) dilution, ab283654 anti-CD68 used at 1 : 100 (4.15 µg/ml) dilution.

Panel A : merged staining of anti-Pentraxin 3 (green; Opal™520), anti-E CD68 (magenta; Opal™570) on mouse lung treated with LPS (1ug/ml 16h)+BFA (1ug/ml 16h) ex vivo.
Panel B : anti-Pentraxin 3 staining stroma cells in mouse lung treated with LPS (1ug/ml 16h)+BFA (1ug/ml 16h) ex vivo.
Panel C : anti-CD68 staining macrophages in mouse lung treated with LPS (1ug/ml 16h)+BFA (1ug/ml 16h) ex vivo.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325639 and ab283654 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.

• WB

Lab

Western blot - Anti-Pentraxin 3/PTX3 antibody [EPR30744-540] (AB325639)

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

Low expression : B16-F10.

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

All lanes:

Western blot - Anti-Pentraxin 3/PTX3 antibody [EPR30744-540] (ab325639)

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 2:

B16-F10 (mouse skin melanoma 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

Observed band size: 42 kDa,36 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-Pentraxin 3/PTX3 antibody [EPR30744-540] (AB325639)

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

Low expression : Neuro-2a.

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

All lanes:

Western blot - Anti-Pentraxin 3/PTX3 antibody [EPR30744-540] (ab325639)

Lane 1:

3T3-L1 (mouse embryonic fibroblast) whole cell lysate at 50 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 50 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: 42 kDa,36 kDa

false

Exposure time: 6s

• WB

Lab

Western blot - Anti-Pentraxin 3/PTX3 antibody [EPR30744-540] (AB325639)

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

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

All lanes:

Western blot - Anti-Pentraxin 3/PTX3 antibody [EPR30744-540] (ab325639)

All lanes:

Mouse skeletal muscle tissue lysate at 50 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: 42 kDa,36 kDa

false

Exposure time: 180s