Skip to main content

S100A9 Antibody

Catalog # NB110-89726 | Novus Biologicals a Bio-Techne Brand
Catalog #
Availability
Size / Price
Qty
Loading...
NB110-89726
NB110-89726SS

Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Product Summary for S100A9 Antibody

Immunogen

Full length human S100A9 protein [Swiss-Prot# P06702] expressed in E. coli.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

16 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for S100A9 Antibody

Western Blot: S100A9 Antibody [NB110-89726] - Cells were transfected with the pCMV6-ENTRY control or pCMV6-ENTRY S100A9 cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-S100A9.
Immunocytochemistry/Immunofluorescence: S100A9 Antibody [NB110-89726] - S100A9 antibody was tested in A431 cells with FITC (green). Nuclei and alpha-tubulin were counterstained with Dapi (blue) and Dylight 550 (red).
Immunohistochemistry-Paraffin: S100A9 Antibody [NB110-89726] - Key molecules of specific signaling pathways are assayed by immunohistochemistry in the colorectum of mice. Immunohistochemistry (200x magnification) of S100a9, in normal control, IgG Ab, and anti-S100a9 Ab-treated colorectal tissues of the colitis-associated cancer mouse (n = 4). Scale bar, 50 um. Staining scores were determined by semi-quantitative optical analysis. Image collected and cropped by CiteAb from the following publication (http://journal.frontiersin.org/article/10.3389/fimmu.2017.01774/full), licensed under a CC-BY license.

Applications for S100A9 Antibody

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:50-1:200

Immunohistochemistry

reported in scientific literature (PMID 25792748)

Immunohistochemistry-Paraffin

reported in scientific literature (PMID 21653680)

Simple Western

1:100

Western Blot

1:1000
Application Notes
In Western blot, a band is seen at approx. 16 kDa.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Published Applications

Read 22 publications using NB110-89726 in the following applications:

Formulation, Preparation, and Storage

Purification

Unpurified

Formulation

Whole antisera

Preservative

0.1% Sodium Azide

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: S100A9

S100A9 (S100 calcium-binding protein A9) is a member of S-100 family (family named so as its members are 100% solubile in ammonium sulfate at normal pH) and it exist as a homodimer or as heterodimer/heterotetramer with S100A8. S100A9 is a calcium binding protein with several post-translation modifications as well as multiple ligands, and when bound to calcium, it binds to arachidonic acid, cytoskeletal elements (e.g. keratin intermediate filaments), RAGE (receptor for advanced glycation endproducts), TLR4, CD36, MMPs, fibronectin, and heparin sulfate glycosaminoglycans for serving diverse biological functions. Localization of S100A9 varies according to cell type/disease state, however, it predominantly localize to cytoplasm, from where it translocates to cytoskeleton/cell membrane upon elevation of intracellular calcium levels. During neutrophil activation or endothelial adhesion of monocytes, it gets secreted via a microtubule-mediated alternative pathway. S100A9 is expressed in normal cell types such as myeloid cells, but its expression levels are increased in conditions of inflammatory diseases (RA, SLE, cystic fibrosis, inflammatory bowel disease, Crohn's disease, Sjogren's syndrome, and progressive systemic sclerosis) as well as in several types of cancers including gastric, esophageal, colon, pancreatic, bladder, ovarian, thyroid, breast and skin cancers.

Long Name

S100 Calcium Binding Protein A9

Alternate Names

Calgranulin B, MRP-14

Entrez Gene IDs

6280 (Human); 20200 (Mouse); 94195 (Rat)

Gene Symbol

S100A9

UniProt

Product Documents for S100A9 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for S100A9 Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Loading...

Protocols

View specific protocols for S100A9 Antibody (NB110-89726):

[[URL:https://www.novusbio.com/products/s100a9-antibody_nb110-89726]][[Captio… Antibody]]
Western Blot Protocol

1. Perform SDS-PAGE (4-12% MES) on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer
apparatus.
3. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% BSA in TBS + Tween, 1 hour at RT.
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
7. Dilute the rabbit anti-S100A9 primary antibody (NB 110-89726) in blocking buffer and incubate 1 hour at room temperature.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers
instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce ECL).

Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.

FAQs for S100A9 Antibody

Showing  1 - 3 of 3 FAQs Showing All
  • Q: We would like to know detail information about immunocytochemistry and immunofluorescence. Could you provide protocol and other image for our reference?

    A: For our ICC testing of this product, our lab tested in A431 cells. This image is visible on our datasheet here: Datasheet. The lab followed the following protocol: Protocol, and used the antibody at a 1:100 dilution.

  • Q: Have any of your S100A9 antibodies been tested for use in neutralization, or blocking assays, on human samples?

    A: Unfortunately at this time we have not tested, or received customer feedback on our S100A9 antibodies for use in blocking or neutralizing assays. If you were planning on testing any of our S100A9 antibodies for neutralizing or blocking capabilities, we would recommend our Innovator’s Reward Program. Under the terms of this program we would be happy to provide a credit for a free vial in exchange for new data on this previously untested or reported application. Please submit this data in the form of an online review. Additional Innovators Reward Program information can be found using this link.

  • Q: We would like to stain paraffin embedded human intestinal tissue for Calprotectin. One question will be which cells express the majority of Calprotectin - invading neutrophils vs epithelium for example. The antibody list you provide is quite extensive. Which antibody would you recommend for our purpose? Which tissue would you recommend as positive control.

    A: Nine of our antibodies to Calprotectin have been validated for IHC-P with human tissue, and you can see all of these products. A number of our IHC images were generated using human spleen samples, and as such this may be a good choice of a positive control tissue. We do sell a human spleen slide product, which is suitable for IHC-P. Unfortunately I do not have an in-depth knowledge of Calprotectin, however the image shown for our antibody with catalogue number NBP1-02826 demonstrates clear staining of neutrophils. The following paper also suggests that the protein is abundant in neutrophils: PMID 11435495.

Showing  1 - 3 of 3 FAQs Showing All
Loading...