Anti-S100A9 抗体

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A9 antibody (AB63818)

IHC image of S100A9 staining in Human Tonsil formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab63818, 0.1 μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-S100A9 antibody (AB63818)

ICC/IF image of ab63818 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab63818, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa, HepG2 and Hek293 cells at 5µg/ml, and in 4% PFA fixed (10 min) HeLa, HepG2, Hek293 and MCF7 cells at 5µg/ml.

• IHC-P

AbReview55709****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100A9 antibody (AB63818)

NBF-fixed mouse spleen tissue section stained for S100A9 using ab63818 at 1/5000 dilution in immunohistochemical analysis. heat mediated antigen retrieval with citrate buffer pH 6 was performed before commencing with the IHC protocol. Goat anti-Rb IgG Alexa Fluor^® 647 was used as the secondary antibody at 1/600 dilution.

This image is courtosy of an abnonymous abreview.

• WB

Project5112****

Western blot - Anti-S100A9 antibody (AB63818)

All lanes:

Western blot - Anti-S100A9 antibody (ab63818) at 1 µg/mL

Lane 1:

Human lymph node tissue lysate - total protein (ab29871) at 10 µg

Lane 2:

Human thymus tissue lysate - total protein (ab30146) at 10 µg

Lane 3:

Human spleen tissue lysate - total protein (ab29699) at 10 µg

Lane 4:

Lung (Human) Tissue Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 13 kDa

Observed band size: 13 kDa

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Exposure time: 5min

• WB

Unknown

Western blot - Anti-S100A9 antibody (AB63818)

All lanes:

Western blot - Anti-S100A9 antibody (ab63818) at 1 µg/mL

All lanes:

Western blot - Recombinant Human S100A9 protein (<a href='/products/proteins-peptides/recombinant-human-s100a9-protein-ab95909'>ab95909</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 13 kDa

Observed band size: 15 kDa

true

Exposure time: 1min

• WB

CiteAb

Western blot - Anti-S100A9 antibody (AB63818)

S100A9 western blot using anti-S100A9 antibody ab63818. Publication image and figure legend from Saul, M. J., Hegewald, A. B., et al., 2019, Front Pharmacol, PubMed 31231226.

ab63818 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 ab63818 please see the product overview.

Validation of proteomics results using Western blot analysis of A) HMGB1, B) S100A9, C) NOX2, D) TLR2, and E) p53 in differentiated MM6 cells with and without knockdown of miR-328 (ΔmiR-328). β-Actin was used as the loading control. The relative changes in ΔmiR-328 samples to control were set as 1 and given as mean + SEM of three independent experiments, except S100A9 expression was analyzed in soluble fraction in four independent experiments, *p < 0.05, **p < 0.01.

false