AF5578

antibody from R&D Systems

Targeting: S100A9 60B8AG, CAGB, CFAG, CGLB, LIAG, MAC387, MIF, MRP14, NIF, P14

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

AF5578

Provider

R&D Systems

Product name

Human S100A9 Antibody

Antibody type

Polyclonal

Description

Antigen Affinity-purified. Detects human S100A9 in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant mouse S100A9 is observed.

Reactivity

Human

Host

Sheep

Conjugate

Unconjugated

Antigen sequence

P06702


Isotype

IgG

Vial size

100 ug

Concentration

LYOPH

Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Western blot

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Human S100A9 by Western Blot. Western blot shows lysates of human peripheral blood mononuclear cells (PBMC), human spleen tissue, human tonsil tissue, and human cartilage tissue. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5578) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for S100A9 at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Western Blot Shows Human S100A9 Specificity by Using Knockout Cell Line. Western blot shows lysates of MDA-MB-468 human breast cancer parental cell line and S100A9 knock out MDA-MB-468 cell line (KO). PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5578) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for S100A9 at approximately 14 kDa (as indicated) in the parental MDA-MB-468 cell line, but is not detectable in knockout MDA-MB-468 cell line. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

S100A9 Specificity is Shown by Immunocytochemistry in Knockout Cell Line. S100A9 was detected in immersion fixed MDA-MB-468 human breast cancer cell line but is not detected in S100A9 knockout (KO) MDA-MB-468 cell line using Sheep Anti-Human S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5578) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

S100A9 in MDA-MB-468 Human Cell Line. S100A9 was detected in immersion fixed MDA-MB-468 human breast cancer cell line using Sheep Anti-Human S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5578) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

S100A9 in Human Cartilage. S100A9 was detected in immersion fixed paraffin-embedded sections of human cartilage using Sheep Anti-Human S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5578) at 1 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of chondrocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Flow cytometry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of S100A9 in Human PBMCs by Flow Cytometry. Human peripheral blood monocytes (PBMC) were stained with Mouse Anti-Human CD14 PE-conjugated Monoclonal Antibody (Catalog # FAB3832P) and either (A) Sheep Anti-Human S100A9 Polyclonal Antibody (Catalog # AF5578) or (B) Sheep IgG Isotype Control (Catalog # 5-001-A) followed by anti-Sheep IgG APC-conjugated Secondary Antibody (Catalog # F0127). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer (Catalog # FC005). View our protocol for Staining Membrane-associated Proteins.