Revision 4

#73425Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-Bond, IHC-P, IF-F, FC-FP

REACTIVITY:

M R

SENSITIVITY:

Endogenous

MW (kDa):

14

Source/Isotype:

Rabbit IgG

UniProt ID:

#P31725

Entrez-Gene Id:

20202

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:800
Immunohistochemistry (Paraffin) 1:800
Immunofluorescence (Frozen) 1:100
Flow Cytometry (Fixed/Permeabilized) 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #74641.

Specificity / Sensitivity

S100A9 (D3U8M) Rabbit mAb recognizes endogenous levels of total S100A9 protein.

Species Reactivity:

Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asn98 of mouse S100A9 protein.

Background

S100A8 and S100A9 are calcium-binding proteins that form a noncovalent heterodimer present in monocytes, neutrophils, macrophages, and some epithelial cells (1,2). S100A8 and S100A9 are secreted by a tubulin-dependent mechanism during inflammatory conditions and have antimicrobial and chemotactic functions (3-5). Extracellular S100A8/S100A9 also induces an inflammatory response in endothelial cells, including induction of proinflammatory chemokines and adhesion molecules and increased vascular permeability (6). S100A8/S100A9 induces and recruits myeloid-derived suppressor cells (MDSC) in tumor-bearing mice (7). MDSC produce additional S100A8/S100A9 themselves, resulting in a positive feedback mechanism that sustains MDSC accumulation (7). S100A8/S100A9 is also highly expressed in psoriatic skin, where it directly upregulates transcription of complement protein C3, which contributes to disease (8). In addition, tumor-infiltrating myeloid cells induce expression of S100A8 and S100A9 in cancer cells, which increases invasiveness and metastasis (9).

  1. Odink, K. et al. (1987) Nature 330, 80-2.
  2. Edgeworth, J. et al. (1991) J Biol Chem 266, 7706-13.
  3. Rammes, A. et al. (1997) J Biol Chem 272, 9496-502.
  4. Steinbakk, M. et al. (1990) Lancet 336, 763-5.
  5. Ryckman, C. et al. (2003) J Immunol 170, 3233-42.
  6. Viemann, D. et al. (2005) Blood 105, 2955-62.
  7. Sinha, P. et al. (2008) J Immunol 181, 4666-75.
  8. Schonthaler, H.B. et al. (2013) Immunity 39, 1171-81.
  9. Lim, S.Y. et al. (2016) Oncogene 35, 5735-45.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 4
#73425

S100A9 (D3U8M) Rabbit mAb

Western Blotting Image 1: S100A9 (D3U8M) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines and tissues using S100A9 (D3U8M) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation Image 1: S100A9 (D3U8M) Rabbit mAb Expand Image
Immunoprecipitation of S100A9 from BaF3 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is S100A9 (D3U8M) Rabbit mAb. Western blot analysis was performed using S100A9 (D3U8M) Rabbit mAb.
Immunohistochemistry Image 1: S100A9 (D3U8M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse ovary using S100A9 (D3U8M) Rabbit mAb performed on the Leica® Bond Rx.
Immunohistochemistry Image 1: S100A9 (D3U8M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded BaF3 cell pellet (left, positive) or Raw 264.7 cell pellet (right, negative) using S100A9 (D3U8M) Rabbit mAb.
Immunohistochemistry Image 2: S100A9 (D3U8M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse spleen using S100A9 (D3U8M) Rabbit mAb.
Immunohistochemistry Image 3: S100A9 (D3U8M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded rat brain using S100A9 (D3U8M) Rabbit mAb.
Immunohistochemistry Image 4: S100A9 (D3U8M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded rat liver using S100A9 (D3U8M) Rabbit mAb.
Immunohistochemistry Image 5: S100A9 (D3U8M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded rat lung using S100A9 (D3U8M) Rabbit mAb.
Immunohistochemistry Image 6: S100A9 (D3U8M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded rat spleen using S100A9 (D3U8M) Rabbit mAb.
Immunohistochemistry Image 7: S100A9 (D3U8M) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse liver using S100A9 (D3U8M) Rabbit mAb.
Immunofluorescence Image 1: S100A9 (D3U8M) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse lung at low (left) and high (right) magnifications using S100A9 (D3U8M) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunofluorescence Image 2: S100A9 (D3U8M) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse spleen at low (left) and high (right) magnifications using S100A9 (D3U8M) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow Cytometry Image 1: S100A9 (D3U8M) Rabbit mAb Expand Image
Flow cytometric analysis of mouse splenocytes, co-stained with anti-mouse CD11b, using S100A9 (D3U8M) Rabbit mAb (right) compared to a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.