Anti-Pentraxin 3/PTX3 antibody [MNB1]

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Pentraxin 3/PTX3 antibody [MNB1] (AB90806)

Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen rat placenta tissue labeling Pentraxin 3/PTX3 with ab90806 at 1/20 dilution followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution. Positive staining on rat placenta was observed. Nuclear counterstain is DAPI (Blue).
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody was ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Pentraxin 3/PTX3 antibody [MNB1] (AB90806)

Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen mouse placenta tissue labeling Pentraxin 3/PTX3 with ab90806 at 1/20 dilution followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution. Positive staining on mouse placenta was observed. Nuclear counterstain is DAPI (Blue).
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody was ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• WB

Lab

Western blot - Anti-Pentraxin 3/PTX3 antibody [MNB1] (AB90806)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1, 2 : 15 seconds; Lane 3 : 3 seconds; Lane 4 : 180 seconds.

All lanes:

Western blot - Anti-Pentraxin 3/PTX3 antibody [MNB1] (ab90806) at 1/1000 dilution

Lane 1:

MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

Mouse placenta tissue lysate at 20 µg

Lane 3:

Rat placenta tissue lysate at 20 µg

Lane 4:

3T3-L1 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 41 kDa

false

• WB

Lab

Western blot - Anti-Pentraxin 3/PTX3 antibody [MNB1] (AB90806)

The expression profile observed is consistent with what has been described in the literature (PMID : 17389238).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Pentraxin 3/PTX3 antibody [MNB1] (ab90806) at 1/1000 dilution

Lane 1:

Untreated HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg

Lane 2:

HUVEC treated with 0.5ug/ml Lipopolysaccharides(LPS) for 4 hours and then added 300 ng/ml Brefeldin A for another 20 hours, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 41 kDa

false

Exposure time: 15s

• WB

CiteAb

Western blot - Anti-Pentraxin 3/PTX3 antibody [MNB1] (AB90806)

Pentraxin 3/PTX3 western blot using anti-Pentraxin 3/PTX3 antibody [MNB1] ab90806. Publication image and figure legend from Scimeca, M., Salustri, A., et al., 2017, Cell Death Dis, PubMed 29022895.

ab90806 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab90806 please see the product overview.

Evaluation of PTX3 in bone head biopsies and osteoblast cell cultures. Immunohistochemical results were evaluated by a semiquantitative approach assigning a score from 0 to 3 according to the number of positive osteoblasts (Table 2). (a) Graph shows immunohistochemical results of PTX3 (OP versus OA P=0.0140; OP versus CTRL p<0.0001; OA versus CTRL p<0.0001). (OP n=25, OA n=25, CTRL n=15) (b and c) Graphs displays the correlation between PTX3 score and BV/TV or Tb.Th values for each OP patient. OP patients are numbered in increasing order of BV/TV or Tb.Th values, respectively. (f and g) OA patients showed high expression of PTX3 both in osteoblasts and bone marrow cells. (h and i) CTRL patients showed expression of PTX3 in almost all osteoblasts and bone marrow cells (g, square). Scale bar represents 200 μm for DFH images and 40 μm for EGI images. (j) Graph shows the results of immunostaining of primary confluent osteoblast cultures (OP n=3, OA n=3, CTRL n=3) (k) Graph shows the presence of micro-HA crystals in primary confluent osteoblast cultures (OP n=3, OA n=3, CTRL n=3). (l and m) Dual-colour immunostaining for PTX3 (red) and RUNX2 (green) on confluent osteoblasts culture derived from CTRL patients. (l) Images show numerous RUNX2-positive osteoblast expressing PTX3. Scale bar represents 50 μm. (m) High magnification image of RUNX2-positive osteoblast expressing PTX3. Scale bar represents 10 μm. (n) Representative images of western blot assay. Western blot analysis displayed remarkable difference in PTX3 expression among the groups. Table reported the ratio between the densitometric values of PTX3 and β-catenin for each experimental group. (OP n=3, OA n=3, CTRL n=3). (o) mRNA expression level of PTX3 relative to GADPH expression in osteoblast primary cultures derived from OP (0.39±0.22), OA (0.72±0.19) and CTRL (1.00±0.11) patients. (OP n=3, OA n=3, CTRL n=3)

false