For Research Use Only. Not For Use In Diagnostic Procedures.

1 Arginase-1 (D4E3M ). Store at -20 C. XP Rabbit mAb Support: +1-978-867-2388 ( ). #93668. Orders: 877-616-2355 ( ). Entrez-Gene ID #383. rev. 01/08/18 UniProt ID #P05089. For Research Use only . Not For Use In Diagnostic Procedures. Applications Species Cross-Reactivity* Molecular Wt. Isotype Storage: Supplied in 10 mM sodium HEPES (pH ), 150. W, IHC-P, IF-IC, IF-F, F mM NaCl, 100 g/ml BSA, 50% glycerol and less than H, M, R 40 kDa Rabbit IgG**. Endogenous sodium azide. Store at 20 C. Do not aliquot the antibody. *Species cross-reactivity is determined by western blot. ** Anti-rabbit secondary antibodies must be used to Background: L-arginine plays a critical role in regulating the detect this antibody. r e liv immune system (1-3). In inflammation, cancer and certain other r e se iv ou Recommended Antibody Dilutions: tl pathological conditions, myeloid cell differentiation is inhibited kDa ra m leading to a heterogeneous population of immature myeloid Western blotting 1:1000.

2 200. cells, known as myeloid-derived suppressor cells (MDSCs). Immunohistochemistry (Paraffin) 1:100 . MDSCs are recruited to sites of cancer-associated inflammation 140 Unmasking buffer: SignalStain Citrate Unmasking Solution and express high levels of arginase-1 (4). Arginase-1 catalyzes (10X) #14746. the final step of the urea cycle converting L-arginine to L- 100 Antibody diluent: SignalStain Antibody Diluent #8112. ornithine and urea (5). Thus MDSCs increase the catabolism of Detection reagent: SignalStain Boost (HRP, Rabbit) #8114. 80. L-arginine resulting in L-arginine depletion in the inflammatory Optimal IHC dilutions determined using SignalStain Boost microenvironment of cancer (4, 6). The reduced availability of IHC Detection Reagent. L-arginine suppresses T-cell proliferation and function and thus 60 Immunohistochemistry (Leica Bond ) 1:400. contributes to tumor progression (4, 6). Arginase-1 is of great 50 Immunofluorescence (IF-IC) 1:50.

3 Interest to researchers looking for a therapeutic target to inhibit Immunofluorescence (IF-F) 1:50. the function of MDSCs in the context of cancer immunotherapy 40 Arginase-1 IF Protocol: Methanol Permeabilization required (7). In addition, Research studies have demonstrated that Flow Cytometry 1:50. Arginase-1 distinguishes primary hepatocellular carcinoma 30. For product specific protocols and a complete listing (HCC) from metastatic tumors in the liver, indicating its value as of recommended companion products please see the a potential biomarker in the diagnosis of HCC (8, 9). product web page at Specificity/Sensitivity: Arginase-1 (D4E3M ) XP Rabbit 20 Background References: mAb recognizes endogenous levels of total arginase-1 protein. This antibody does not cross-react with arginase-2. (1) Albina, et al. (1989) J Exp Med 169, 1021-9. Western blot analysis of extracts from mouse and rat livers using (2) Mills, (2001) Crit Rev Immunol 21, 399-425.

4 Source/Purification: Monoclonal antibody is produced by Arginase-1 (D4E3M ) XP Rabbit mAb. immunizing animals with a synthetic peptide corresponding to (3) Rodriguez, et al. (2004) Cancer Res 64, 5839-49. residues surrounding Val47 of human arginase-1 protein. (4) Gabrilovich, and Nagaraj, S. (2009) Nat Rev Immunol 9, 162-74. (5) Wu, G. and Morris, (1998) Biochem J 336 ( Pt 1), 1-17. (6) Raber, P. et al. (2012) Immunol Invest 41, 614-34. (7) Wesolowski, R. et al. (2013) J Immunother Cancer 1, 10. (8) Sang, W. et al. (2015) Tumour Biol 36, 3881-6. (9) Geramizadeh, B. and Seirfar, N. (2015) Hepat Mon 15, e30336. Alexa Fluor is a registered trademark of Life Technologies Corporation. BOND is a trademark of Leica Biosystems Melbourne Pty. Ltd. No affiliation or sponsorship between CST and Leica Microsystems IR GmbH or Leica Biosystems Melbourne Pty. Ltd is implied. DRAQ5 is a registered trademark of Biostatus Limited. DyLight is a trademark of Thermo Fisher Scientific Inc.

5 And its subsidiaries. Immunohistochemical analysis of paraffin-embedded normal human Immunohistochemical analysis of paraffin-embedded human hepato- LEICA is a registered trademark of Leica Microsystems IR GmbH. liver using Arginase-1 (D4E3M ) XP Rabbit mAb. cellular carcinoma using Arginase-1 (D4E3M ) XP Rabbit mAb. Tween is a registered trademark of ICI Americas, Inc. Thank you for your recent purchase. If you would like to provide a review visit IMPORTANT: For western blots, incubate membrane with diluted antibody in w/v 5% nonfat dry milk, 1X TBS, Tween 20 at 4 C with gentle shaking, overnight. 2015 Cell Signaling Technology, Inc. D4E3M, XP, SignalStain, and Cell Signaling Technology are trademarks of Cell Signaling Technology, Inc. Applications: W Western IP Immunoprecipitation IHC Immunohistochemistry ChIP Chromatin Immunoprecipitation IF Immunofluorescence F Flow cytometry E-P ELISA-Peptide Species Cross-Reactivity: H human M mouse R rat Hm hamster Mk monkey Mi mink C chicken Dm D.

6 Melanogaster X Xenopus Z zebrafish B bovine Dg dog Pg pig Sc S. cerevisiae Ce C. elegans Hr Horse All all species expected Species enclosed in parentheses are predicted to react based on 100% homology. mouse small intestine mouse liver kDa 200. 140. 100. 80. 60. 50. Immunohistochemical analysis of paraffin-embedded human Immunohistochemical analysis of paraffin-embedded mouse 40 Arginase-1. lung carcinoma using Arginase-1 (D4E3M ) XP Rabbit mAb. liver using Arginase-1 (D4E3M ) XP Rabbit mAb. 30. mouse liver mouse small intestine 20. 50 -Actin 40 (D6A8). 30. Western blot analysis of extracts from mouse liver and mouse small intestine using Arginase-1 (D4E3M ) XP Rabbit mAb (upper) or -Actin (D6A8) Rabbit mAb #8457 (lower). Confocal immunofluorescent analysis of mouse liver (positive; left) or small intestine (negative; right). using Arginase-1 (D4E3M ) XP Rabbit mAb (green). Blue pseudocolor = DRAQ5 #4084 (fluorescent DNA dye). IL-4/cAMP LPS/IFNg Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Arginase-1 (D4E3M ) XP Rabbit mAb performed on the Leica Bond Rx.

7 Confocal immunofluorescent analysis of mouse primary bone marrow-derived macrophages (BMDMs) using Arginase-1 (D4E3M ) XP Rabbit mAb (green). BMDMs were differentiated with M-CSF (20 ng/ml, 7 days). and activated with either IL-4/cAMP (20 ng/ml, mM, 24 hours; left) or LPS/IFN (50 ng/ml, 20 ng/ml, 24. Events hours; right). Red = Propidium Iodide (PI)/RNase Staining Solution #4087. Arginase-1. Flow cytometric analysis of human whole blood using Arginase-1. (D4E3M ) XP Rabbit mAb (solid line) compared to concentra- tion-matched Rabbit (DA1E) mAb IgG XP Isotype Control #3900. (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488 Conjugate) #4412 was used as a secondary antibody. Analysis was performed on cells in the granulocyte gate. Thank you for your recent purchase. If you would like to provide a review visit 2016 Cell Signaling Technology, Inc.