Cathepsin D Monoclonal antibody

• Featured Product
• KD/KO Validated

Cathepsin D Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

human and More (1)

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

8

CloneNo.

2F6F7

Cat no : 66534-1-Ig

Synonyms

CTSD, CPSD, CLN10, Cathepsin D light chain, Cathepsin D heavy chain

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Validation Data Gallery

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  WB analysis using 66534-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66534-1-Ig (Cathepsin D antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of MCF-7 using 66534-1-Ig

  MCF-7 cells were subjected to SDS PAGE followed by western blot with 66534-1-Ig (Cathepsin D antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  IHC staining of human liver cancer using 66534-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66534-1-Ig (Cathepsin D antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human liver cancer using 66534-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66534-1-Ig (Cathepsin D antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 66534-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66534-1-Ig (Cathepsin D antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 66534-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66534-1-Ig (Cathepsin D antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver cancer using 66534-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66534-1-Ig (Cathepsin D antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver cancer using 66534-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66534-1-Ig (Cathepsin D antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HepG2 using 66534-1-Ig

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using Cathepsin D antibody (66534-1-Ig, Clone: 2F6F7 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

• 

  FC experiment of HepG2 using 66534-1-Ig

  1X10^6 HepG2 cells were intracellularly stained with 0.4 ug Anti-Human Cathepsin D (66534-1-Ig, Clone:2F6F7) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG2b Isotype Control (MPC-11) (65128-1-Ig, Clone: MPC-11) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: Saos-2 cells, MCF-7 cells, T-47D cells, HepG2 cells
• Positive IHC detected in: human liver cancer tissue, human breast cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 1 publications below
• WB: See 6 publications below
• IHC: See 1 publications below
• IF: See 2 publications below

Product Information

66534-1-Ig targets Cathepsin D in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human, rat
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: Cathepsin D fusion protein Ag15254
• Full Name: cathepsin D
• Calculated Molecular Weight: 412 aa, 45 kDa
• Observed Molecular Weight: 32 kDa, 48 kDa, 52 kDa
• GenBank Accession Number: BC016320
• Gene Symbol: Cathepsin D
• Gene ID (NCBI): 1509
• RRID: AB_2881897
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

CTSD (Cathepsin D) is also named as CPSD, belongs to the peptidase A1 family. It is ubiquitously expressed and is involved in proteolytic degradation, cell invasion, and apoptosis. Human CTSD is synthesized as a 52-kDa precursor that is converted into an active 48-kDa single-chain intermediate in the endosomes, and then into a fully active mature form, composed of a 34-kDa heavy chain and a 14-kDa light chain, in the lysosomes. It is a lysosomal acid protease found in neutrophils and monocytes and involved in the pathogenesis of several diseases such as breast cancer and possibly Alzheimer disease. (PMID: 27114232, PMID: 30717773, PMID: 30051532)

Publications

Species	Application	Title
human	IHC	Gastroenterology; Proteomic characterization identifies clinically relevant subgroups of gastrointestinal stromal tumors; Authors - Mingjun Sun
rat	WB,IF	Life (Basel); Melibiose Confers a Neuroprotection against Cerebral Ischemia/Reperfusion Injury by Ameliorating Autophagy Flux via Facilitation of TFEB Nuclear Translocation in Neurons.; Authors - Zhiyuan Wu
	WB	Cell Rep; Intracellular complement C5a/C5aR1 stabilizes β-catenin to promote colorectal tumorigenesis.; Authors - Peipei Ding ; KD Validated
human	IF	FEBS Lett; Senile plaques in Alzheimer's disease arise from Aβ- and Cathepsin D-enriched mixtures leaking out during intravascular haemolysis and microaneurysm rupture; Authors - Hualin Fu
human,rat	WB	Ecotoxicol Environ Saf; The inhibition of TRPML1/TFEB leads to lysosomal biogenesis disorder, contributes to developmental fluoride neurotoxicity; Authors - Jingjing Zhang
rat	WB	Ecotoxicol Environ Saf; TFE3-mediated impairment of lysosomal biogenesis and defective autophagy contribute to fluoride-induced hepatotoxicity; Authors - Zeyu Hu