Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot, Neutralization, Simple Western, Immunoprecipitation

Cited:

Western Blot, Array Development, ELISA Development

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Cystatin C Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Cystatin C
Ser27-Ala146
Accession # P01034

Specificity

Detects human Cystatin C in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 50% cross-reactivity with recombinant mouse Cystatin C is observed and less than 5% cross-reactivity with recombinant human (rh) Fetuin A, rhFetuin B, rhCystatin A, rhCystatin B, rhCystatin S, and rhCystatin E/M is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human Cystatin C Antibody

Detection of Human Cystatin C antibody by Western Blot.

Detection of Human Cystatin C by Western Blot.

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, human liver tissue, and human serum. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human Cystatin C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1196) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). 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.
Detection of Human Cystatin C antibody by Western Blot.

Detection of Human Cystatin C by Western Blot.

Western blot shows lysates of human placenta tissue and PC-3 human prostate cancer cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human Cystatin C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1196) 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 antibody in Human Breast by Immunohistochemistry (IHC-P).

Cystatin C in Human Breast.

Cystatin C was detected in immersion fixed paraffin-embedded sections of human breast using 5 µg/mL Goat Anti-Human Cystatin C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1196) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the cytoplasm in cells of terminal ductules. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Cystatin C antibody by Simple WesternTM.

Detection of Human Cystatin C by Simple WesternTM.

Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Cystatin C at approximately 20 kDa (as indicated) using 10 µg/mL of Goat Anti-Human Cystatin C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1196). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of Human Cystatin C by Western Blot

Detection of Human Cystatin C by Western Blot

Cystatin E/M and C secretions from various cell lines: Common laboratory non-melanoma and established laboratory melanoma cell lines (top), and primary and metastatic melanoma cell lines established from patients, as well as skin control (bottom). (A) Equal amounts of serum free media were collected from 5 × 105 cells 48 h after changing from ordinary growth media and secreted proteins were concentrated by TCA-precipitation and subjected to SDS-PAGE and immunoblotting. The filters were stained with a cystatin E/M-specific (upper panels) or a cystatin C-specific (lower panel) antibody, respectively. (B) Inhibitory activity against legumain was measured in the conditioned media as residual legumain activity. A partially purified legumain fraction from rat kidney was mixed with conditioned media and the ability to cleave the substrate Z-Ala-Ala-Asn-NHMec was measured by fluorometry. Control bar (100%) represents activity in the rat legumain fraction without addition of conditioned media. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/20074384), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Cystatin C Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human breast

Immunoprecipitation

25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant Human Cystatin C (Catalog # 1196-PI), see our available Western blot detection antibodies

Simple Western

10 µg/mL
Sample: Human hepatocellular carcinoma cell line

Western Blot

0.25-2 µg/mL
Sample: HepG2 human hepatocellular carcinoma cell line, human liver tissue, human serum, human placenta tissue and PC‑3 human prostate cancer cell line

Neutralization

Measured by its ability to neutralize Recombinant Human Cystatin C (0.64 µg/mL, Catalog # 1196-PI) inhibition of Papain (0.1 µg/mL) cleavage of the fluorogenic peptide substrate Z-FR-AMC (100 µM, Catalog # ES009). The Neutralization Dose (ND50) is typically 12-112 µg/mL.

Reviewed Applications

Read 2 reviews rated 4.5 using AF1196 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Cystatin C

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). It is ubiquitous in human tissues and body fluids. A point mutation in the gene coding for the 120 amino acid mature Cystatin C causes a hereditary form of amyloid angiopathy in which the protein variant (Leu68 to Gln) is deposited in the cerebral arteries, leading to fatal cerebral hemorrhage (4). Cystatin C may have additional clinical applications. For example, it is a good marker for glomerular filtration rate (5).

References

  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. Abrahamson, M. et al. (1992) Hum. Genet. 89:377.
  5. Laterza, O.F. et al. (2002) Clin. Chem. 48:699.

Alternate Names

ARMD11, CST3, Gamma-trace, Neuroendocrine basic polypeptide, Post-gamma-globulin

Entrez Gene IDs

1471 (Human); 13010 (Mouse); 25307 (Rat)

Gene Symbol

CST3

UniProt

P01034

Additional Cystatin C Products

Product Documents for Human Cystatin C Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Cystatin C Antibody

For research use only

Citations for Human Cystatin C Antibody

Customer Reviews for Human Cystatin C Antibody (2)

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  • Human Cystatin C Antibody
    Name: Estefania Torrejon
    Application: Western Blot
    Sample Tested: HepG2 human hepatocellular carcinoma cell line
    Species: Human
    Verified Customer | Posted 05/17/2022
    Good signal at 40 ug Incubated ON at 4 C Dilution of 1:500
    Human Cystatin C Antibody AF1196
  • Human Cystatin C Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Recombinant protein
    Species: Human
    Verified Customer | Posted 12/17/2020
    Human Cystatin C Antibody AF1196

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Protocols

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