Cathepsin D Antibody [Unconjugated]

Western blot shows lysate of mouse intestine tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse Cathepsin D Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1029) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for Cathepsin D at approximately 28 and 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Cathepsin D was detected in perfusion fixed frozen sections of mouse intestine using 15 µg/mL Goat Anti-Mouse Cathepsin D Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1029) 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 epithelial cells in intestinal villi. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Cathepsin D was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line using Goat Anti-Mouse Cathepsin D Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1029) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to lysosomes. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

• Reactivity: Mu
• Applications: WB, IHC, IP, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Cathepsin D Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse Cathepsin D
  Ile21-Leu410
  Accession # Q3UCD9
• Specificity: Detects mouse Cathepsin D in direct ELISAs and Western blots. In direct ELISAs, approximately 15% cross-reactivity with recombinant human Cathepsin D is observed and less than 1% cross-reactivity with recombinant mouse Cathepsin E is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CTSD

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Immunoprecipitation 25 ug/mL
  • Immunocytochemistry 5-15 ug/mL

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 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 Cathepsin D Antibody [Unconjugated]

• cathepsin D (lysosomal aspartyl protease)
• Cathepsin D
• CPSD
• CTSD
• EC 3.4.23
• EC 3.4.23.5
• lysosomal aspartyl peptidase
• lysosomal aspartyl protease
• MGC2311
• neuronal 10

Background

Cathepsin D is a lysosomal aspartic protease of the pepsin family (4). Mouse Cathepsin D is synthesized as a precursor protein, consisting of a signal peptide (residues 1-20), a propeptide (residues 21-64), and a mature chain (residues 65-410) (1-3). It is expressed in most cells and over-expressed in breast cancer cells (5). It is a major enzyme in protein degradation in lysosomes, and also involved in the presentation of antigenic peptides. Mice deficient in this enzyme showed a progressive atrophy of the intestinal mucosa, a massive destruction of lymphoid organs, and a profound neuronal ceroid lipofucinosis, indicating that Cathepsin D is essential for proteolysis of proteins regulating cell growth and tissue homeostasis (6). Cathepsin D secreted from human prostate carcinoma cells is responsible for the generation of angiostatin, a potent endogeneous inhibitor of angiogenesis (6).

1. Diedrich, et al. (1990) Nucl. Acid Res. 18:7184.
2. Grusby, et al. (1990) Nucl. Acid Res. 18:4008. 
3. Hetman, et al. (1994) DNA Cell Biol. 13:419.
4. Conner, (2004) in Handbook of Proteolytic Enzymes (Barrett, et al. eds) Elsevier Academic Press, San Diego, p. 43.
5. Rochefort, et al. (2000) Clin. Chim. Acta. 291:157.
6. Tsukuba, et al. (2000) Mol. Cells 10:601.

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 Cathepsin D Antibody (AF1029)(13)

Publications using AF1029

• Q Shao, M Yang, C Liang, L Ma, W Zhang, Z Jiang, J Luo, JK Lee, C Liang, JF Chen C9orf72 and smcr8 mutant mice reveal MTORC1 activation due to impaired lysosomal degradation and exocytosis Autophagy, 2019;0(0):1-16. 2019 [PMID: 31847700] (ICC, Mouse)
• Y Atiskova, S Bartsch, T Danyukova, E Becker, C Hagel, S Storch, U Bartsch Mice deficient in the lysosomal enzyme palmitoyl-protein thioesterase 1 (PPT1) display a complex retinal phenotype Sci Rep, 2019;9(1):14185. 2019 [PMID: 31578378] (Western Blot, Mouse)
• A Kishimoto, S Kimura, J Nio-Kobaya, H Takahashi-, AM Park, T Iwanaga Histochemical characteristics of regressing vessels in the hyaloid vascular system of neonatal mice: Novel implication for vascular atrophy Exp. Eye Res., 2018;172(0):1-9. 2018 [PMID: 29596849] (IHC, Mouse)
• Plasma cathepsin D correlates with histological classifications of fatty liver disease in adults and responds to intervention Sci Rep, 2016;6(0):38278. 2016 [PMID: 27922112] (IHC, Human)
• Kenneth M Murphy RAB43 facilitates cross-presentation of cell-associated antigens by CD8?+ dendritic cells J. Exp. Med., 2016;0(0):. 2016 [PMID: 27899443] (Western Blot, Mouse)
• A C9ORF72/SMCR8-containing complex regulates ULK1 and plays a dual role in autophagy Sci Adv, 2016;2(9):e1601167. 2016 [PMID: 27617292] (Western Blot, Mouse)
• Jensen, Thomas B, Cheema, Muhammad, Szymiczek, Agata, Damkier, Helle Ha, Praetorius, Jeppe Renal type a intercalated cells contain albumin in organelles with aldosterone-regulated abundance. PLoS ONE, 2015;10(4):e0124902. 2015 [PMID: 25874770] (IHC, Mouse)
• Kern U, Wischnewski V, Biniossek M, Schilling O, Reinheckel T Lysosomal protein turnover contributes to the acquisition of TGFbeta-1 induced invasive properties of mammary cancer cells. Mol Cancer, 2015;14(0):39. 2015 [PMID: 25744631] (Western Blot, Mouse)
• Lee J, Jin H, Park M, Kim B, Lee P, Nakauchi H, Carter J, He X, Schuchman E, Bae J Acid sphingomyelinase modulates the autophagic process by controlling lysosomal biogenesis in Alzheimer's disease. J Exp Med, 2014;211(8):1551-70. 2014 [PMID: 25049335] (Western Blot, Mouse)
• Lai, N Chin, Gao, Mei Hua, Tang, Eric, Tang, Ruoying, Guo, Tracy, Dalton, Nancy D, Deng, Aihua, Tang, Tong Pressure overload-induced cardiac remodeling and dysfunction in the absence of interleukin 6 in mice. Lab Invest, 2012;92(11):1518-26. 2012 [PMID: 22825686] (Western Blot, Mouse)
• Zeeuwen PL, van Vlijmen-Willems IM, Cheng T The cystatin M/E-cathepsin L balance is essential for tissue homeostasis in epidermis, hair follicles, and cornea. FASEB J., 2010;24(10):3744-55. 2010 [PMID: 20495178] (Western Blot, Mouse)
• Schahs P, Weidinger P, Probst OC, Svoboda B, Stadlmann J, Beug H, Waerner T, Mach L Cellular repressor of E1A-stimulated genes is a bona fide lysosomal protein which undergoes proteolytic maturation during its biosynthesis. Exp. Cell Res., 2008;314(16):3036-47. 2008 [PMID: 18621046] (Western Blot, Mouse)
• Uppington H, Menager N, Boross P, Wood J, Sheppard M, Verbeek S, Mastroeni P Effect of immune serum and role of individual Fcgamma receptors on the intracellular distribution and survival of Salmonella enterica serovar Typhimurium in murine macrophages. Immunology, 2006;119(2):147-58. 2006 [PMID: 16836651] (ICC, Mouse)

Bioinformatics

• Gene Symbol: CTSD
• Uniprot:
  • Q3UCD9()