Cystatin C Antibody [Unconjugated]

Western blot shows lysates of mouse plasma and RAW 264.7 mouse monocyte/macrophage cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse Cystatin C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1238) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Cystatin C at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Cystatin C was detected in immersion fixed frozen sections of mouse spleen using Mouse Cystatin C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1238) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Simple Western lane view shows lysates of RAW 264.7 mouse monocyte/macrophage cell line, loaded at 0.2 mg/mL. A specific band was detected for Cystatin C at approximately 19 kDa (as indicated) using 10 µg/mL of Goat Anti-Mouse Cystatin C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1238) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reactivity: Mu
• Applications: WB, Simple Western, IHC, IP, ELISA(Cap)
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Cystatin C Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse Cystatin C (R & D Systems, Catalog # 1238-PI)
  Ala21-Ala140
  Accession # P21460
• Specificity: Detects mouse and human Cystatin C in direct ELISAs. In direct ELISAs, less than 10% cross-reactivity with recombinant human Cystatin D is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CST3
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • ELISA Capture (Matched Antibody Pair) 0.2-0.8 ug/mL
  • ELISA Detection (Matched Antibody Pair) 0.1-0.4 ug/mL
  • ELISA Standard (Matched Pair)
  • Immunohistochemistry 5-15 ug/mL
  • Immunoprecipitation 25 ug/mL
  • Simple Western 10 ug/mL
  • Western Blot 1 ug/mL
• Application Notes: ELISA Capture:Mouse Cystatin C Antibody (Catalog # AF1238)
  ELISA Detection:Mouse Cystatin C Biotinylated Antibody (Catalog # BAF1238)
  Standard: Recombinant Mouse Cystatin C (Catalog # 1238-PI)

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Cystatin C Antibody [Unconjugated]

• ARMD11
• bA218C14.4 (cystatin C)
• CST3
• cystatin 3
• cystatin C (amyloid angiopathy and cerebral hemorrhage)
• Cystatin C
• cystatin-3
• cystatin-C
• Gamma-trace
• MGC117328
• Neuroendocrine basic polypeptide
• Post-gamma-globulin

Background

Cystatin C is a member of family 2 of the cystatin superfamily (1). It is involved in processes such as tumor invasion and metastasis, inflammation and some neurological diseases. It inhibits many cysteine proteases such as papain and cathepsins B, H, K, L and S (2, 3). All mouse tissues analyzed expressed Cystatin C, with relative levels similar to those of rat and human tissues. For all three species, brain and liver had the highest and lowest levels of Cystatin C, respectively, whereas kidney, spleen and muscle had the levels in between (4). The high degree of similarity in distribution and functional properties for mouse, rat and human Cystatin C indicates that a murine model should be relevant for studies of the human disease, hereditary Cystatin C amyloid angiopathy (4).

1. Reed, C.H. (2000) British J. Biomed. Sci. 57:323.
2. Janowski, R. et al. (2001) Nat. Struct. Biol. 8:316.
3. Abrahamson, M. (1994) Methods Enzymol. 244:685.
4. Hakansson, K. et al. (1996) Comp. Biochem. Physiol. 114B:303.

Limitations

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.

Publications for Cystatin C Antibody (AF1238)(6)

Publications using AF1238

• Xiaoxiao Zhang, Xinyue Liu, Guowei Su, Miaomiao Li, Jian Liu, Chunyu Wang et al. pH-dependent and dynamic interactions of cystatin C with heparan sulfate Communications Biology 2/12/2021 [PMID: 33580179]
• T Masuda, R Sankowski, O Staszewski, C Böttcher, L Amann, C Scheiwe, S Nessler, P Kunz, G van Loo, VA Coenen, PC Reinacher, A Michel, U Sure, R Gold, J Priller, C Stadelmann, M Prinz Spatial and temporal heterogeneity of mouse and human microglia at single-cell resolution Nature, 2019-02-13;0(0):. 2019-02-13 [PMID: 30760929] (IHC-P, Mouse)
• D Jensen, C Kierulf-La, MLV Kristensen, R Nørregaard, K Weyer, R Nielsen, EI Christense, H Birn Megalin dependent urinary cystatin C excretion in ischemic kidney injury in rats PLoS ONE, 2017-06-02;12(6):e0178796. 2017-06-02 [PMID: 28575050] (Western Blot, Mouse)
• Yikun Li, Eric B. Dammer, Xiaohui Zhang-Brotzge, Scott Chen, Duc M. Duong, Nicholas T. Seyfried et al. Osteopontin Is a Blood Biomarker for Microglial Activation and Brain Injury in Experimental Hypoxic-Ischemic Encephalopathy eNeuro 1/4/2017 [PMID: 28101531]
• Zhang X, Wu Z, Hayashi Y, Okada R, Nakanishi H Peripheral role of cathepsin S in Th1 cell-dependent transition of nerve injury-induced acute pain to a chronic pain state. J Neurosci, 2014-02-19;34(8):3013-22. 2014-02-19 [PMID: 24553941] (IHC-Fr, Western Blot, Mouse)
• Dieterle F, Perentes E, Cordier A, Roth DR, Verdes P, Grenet O, Pantano S, Moulin P, Wahl D, Mahl A, End P, Staedtler F, Legay F, Carl K, Laurie D, Chibout SD, Vonderscher J, Maurer G Urinary clusterin, cystatin C, beta2-microglobulin and total protein as markers to detect drug-induced kidney injury. Nat. Biotechnol., 2010-05-01;28(5):463-9. 2010-05-01 [PMID: 20458316] (ELISA Development, Human)

Bioinformatics

• Gene Symbol: CST3
• Uniprot:
  • P21460()