Overview

  • Product name
    Anti-Cathepsin D antibody [CTD-19]
    See all Cathepsin D primary antibodies
  • Description
    Mouse monoclonal [CTD-19] to Cathepsin D
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, ELISA, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Full length native protein (purified) (Human liver).

  • Positive control
    • Human breast carcinoma tissue or cell line extract.

Properties

Applications

Our Abpromise guarantee covers the use of ab6313 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 52 kDa.
ELISA Use at an assay dependent concentration.
IHC-P 1/200.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration. PubMed: 24833013

Target

  • Function
    Acid protease active in intracellular protein breakdown. Involved in the pathogenesis of several diseases such as breast cancer and possibly Alzheimer disease.
  • Tissue specificity
    Expressed in the aorta extrcellular space (at protein level).
  • Involvement in disease
    Ceroid lipofuscinosis, neuronal, 10
  • Sequence similarities
    Belongs to the peptidase A1 family.
    Contains 1 peptidase A1 domain.
  • Post-translational
    modifications
    N- and O-glycosylated.
  • Cellular localization
    Lysosome. Melanosome. Secreted, extracellular space. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. In aortic samples, detected as an extracellular protein loosely bound to the matrix (PubMed:20551380).
  • Information by UniProt
  • Database links
  • Alternative names
    • CatD antibody
    • CATD_HUMAN antibody
    • Cathepsin D antibody
    • Cathepsin D heavy chain antibody
    • CD antibody
    • Ceroid lipofuscinosis neuronal 10 antibody
    • CLN10 antibody
    • CPSD antibody
    • ctsd antibody
    • Epididymis secretory sperm binding protein Li 130P antibody
    • HEL S 130P antibody
    • Lysosomal aspartyl peptidase antibody
    • Lysosomal aspartyl protease antibody
    • MGC2311 antibody
    see all

Images

  • Anti-Cathepsin D antibody [CTD-19] (ab6313) at 1/1000 dilution + MDA MB 231 whole cell lysate at 20 µg

    Secondary
    HRP-conjugated monoclonal sheep anti-mouse IgG at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 28,48,50 kDa
    why is the actual band size different from the predicted?


    Exposure time: 2 minutes


    Blocked with 5% milk for 1 hour at 22°C.

    Incubated with the primary antibody for 1 hour at 22°C in 5% TBS-T.

    See Abreview

  • ab6313 staining Cathepsin D in DU145 human prostate cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 pH 7.4 and blocked with 5% BSA at room temperature for 20 minutes. Samples were incubated with primary antibody (1/200 in PBS) for 1 hour. A CF488A-conjugated goat anti-mouse IgG (H+L) polyclonal was used as the secondary antibody (1/500).

    See Abreview

  • ab6313 staining Cathepsin D in HepaRG cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with Triton X-100 in PBS and blocked with 1% milk at room temperature for 20 minutes. Samples were incubated with primary antibody (1µg/ml in 1% milk) for 30 minutes. An undiluted Alexa Fluor® 594-conjugated goat anti-mouse IgG  polyclonal was used as the secondary antibody.

    See Abreview

  • Anti-Cathepsin D antibody [CTD-19] (ab6313) at 1/4000 dilution + MCF7 Human breast carcinoma cells

References

This product has been referenced in:
  • Magalhaes J  et al. Effects of ambroxol on the autophagy-lysosome pathway and mitochondria in primary cortical neurons. Sci Rep 8:1385 (2018). WB ; Mouse . Read more (PubMed: 29362387) »
  • Duta-Mare M  et al. Lysosomal acid lipase regulates fatty acid channeling in brown adipose tissue to maintain thermogenesis. Biochim Biophys Acta 1863:467-478 (2018). IHC-P ; Mouse . Read more (PubMed: 29374543) »
See all 34 Publications for this product

Customer reviews and Q&As

1-10 of 23 Abreviews or Q&A

Application
Western blot
Sample
Human Tissue lysate - whole (CSF)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
50 µg
Specification
CSF
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 11 2015

Application
Western blot
Sample
Mouse Tissue lysate - whole (brain tissue)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
50 µg
Specification
brain tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 11 2015

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Non-reduced Denaturing (10)
Sample
Human Cell lysate - other (Glioblastoma cell lines)
Specification
Glioblastoma cell lines
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Mr. Achilleas Mitrakas

Verified customer

Submitted Apr 25 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C
Sample
Human Cell (DU145 Human prostate)
Specification
DU145 Human prostate
Permeabilization
Yes - 0.1% v/v Triton X-100 pH 7.4 for 5 min at RT
Fixative
Paraformaldehyde

Dr. Dimitra Kalamida

Verified customer

Submitted Nov 21 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (adipocyte tissue)
Loading amount
35 µg
Specification
adipocyte tissue
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 18 2012

Answer

Vielen Dank für Ihre Anfrage die Herr Hryca uns in den technischen Dienst weitergeleitet hat.

Ich habe geschaut, und wir haben viele möglichen positiven Kontrollen für Sie. Können Sie mir bitte noch sagen, welche Art von positive Kontrolle Sie benötigen? Ist es ein Lysat für Western blot oder ein Gewebeschnitt für Immunohistochemie? Haben Sie auch eine Präferenz für eine Spezies (Maus, human)?

Hier mal eine Aufstellung von möglichen positiven Kontrollen. Ich freue mich wieder von Ihnen zu hören, um Ihnen genaure Vorschläge machen zu können.




cell lines Slide
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=2105 Rabbit polyclonal to IL1 beta LPS treated RAW 264.7 cells, Alveolar macrophages (MHS cell line) liver, diseased, inflammed tissue, monocytes, macrophages
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=6313 Mouse monoclonal [CTD-19] to Cathepsin D SKBR3, MCF7 Human liver tissue.
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=6535 Mouse monoclonal [BRM-22] to Hsp70 Hela, 293T,MCF7, Jurkat, HepG2, A431, HCT116, PC3, ... liver, brain, testes, heart, skeletal muscle
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=8219 Mouse monoclonal [MEM-259] to CD63 HT1080, HepG2, OVCAR-3, HL60 and A375 hemapoietic ells, tissue macrophages, platelet, human melanoma tissue, monocytes, macrophages, endothelial cells, SM cells, brain white matter, peripheral nerves
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=25631 Mouse monoclonal [H4B4] to LAMP2 - Lysosome Marker HepG2, 3T3, HeLa, RAW264.7, ... placenta, lung, liver
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=50533 Mouse monoclonal [Rab7-117] to RAB7 HepG2, B16, SK-MEL-28, 3T3, Human brain whole tissue, mouse neuroblastoma
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=68340 Mouse monoclonal to Rab9 Hela, A172, 3T3, 293, PC-12. COS-7, THP-1 tonsil, spleen, prostate cancer, stomach tissu
http://ops.adminsite.com/admin/data/fs_ab.cfm?intAbID=72907 Rabbit polyclonal to Rab5b Hela, Jurkat, HEK293, HepG2 Human Placenta Tissue, Human Brain Tissue.

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Question

1) Abcam product code ab 6313

2) Abcam order reference number or product batch number
Purchase order No.1129425/11200/SK, Order No.1038011, Lot No. GR43577
3) Description of the problem
Very weak non-specific signal by IF
4) Sample preparation:
Species : Human (Hela cell), Rat (PC12 cell)
Type of sample: cells in culture
Sample preparation : See below
Positive control
Negative control : No primary Ab
5) Fixation step
Yes
If yes: Fixative agent and concentration : 4% PFA or MeOH
Fixation time :10 min or 60 min
Fixation temperature : R/T
6) Antigen retrieval method : No
7) Permeabilization method:
Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers? :
permeabilization step : 0.1% Triton X100/PBS or 0.05% Saponin/PBS for 10min
permeabilizing agent in any dilution buffers : same with above
Permeabilizing agent and concentration: 0.1% Triton X100/PBS or 0.05% Saponin/PBS
8) Blocking agent (eg BSA, serum…):
Concentration : 1% Goat serum
Blocking time : 10min
Blocking temperature : R/T
9) Endogenous peroxidases blocked?
Endogenous biotins blocked?
Not used peroxidase system.
10) Primary antibody (If more than one was used, describe in “additional notes”) :
Concentration or dilution : 1/400-1/1,000
Diluent buffer : 1% Goat serum + 0.1% Triton X100/PBS or 0.05% Saponin/PBS
Incubation time : 1 hr
11) Secondary antibody: anti-mouse IgG Alexa488
Species : Goat
Reacts against : Mouse
Concentration or dilution : 1/400
Diluent buffer : 1% Goat serum + 0.1% Triton X100/PBS or 0.05% Saponin/PBS
Incubation time : 1 hr
Fluorochrome or enzyme conjugate : Fluorochrome
12) Washing after primary and secondary antibodies:
Buffer : 0.1% Triton X100/PBS or 0.05% Saponin/PBS
Number of washes : 5 min x3
13) Detection method : Fluorescent microscopy
14) How many times have you run this staining? 3 times
Do you obtain the same results every time? Yes
What steps have you altered to try and optimize the use of this antibody?
Concentration of primary Ab, Detergent, Fixation
Document attachment: Attaching images of your IHC is strongly recommended and can greatly speed up our investigation of your problem.

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Answer

Thank you for providing your protocol details.


I have looked at the protocols used and everything seems fine to me. Therefore, I am more than happy to arrange for one vial of ab21679 Anti-Clathrin heavy chain antibody as a free of charge replacement to you. We currently have stock in the UK office and the product should reach you within a few days (order # 1135414).


If there is anything else that I can help you with, please do not hesitate to contact me.

Have a nice day!

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Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.


Unfortunately, the attachment didn't reach us. This shouldn't be a problem if you mention the PO number or delivery date in the questionnaire.


In addition, am I assuming correctly that you are Japanese? If this is the case, than I am delighted to tell you that we can provide technical support in Japanese as well. Just send your reply to the email address below. Otherwise, just reply to my email as you normally would.

mailto:technical@abcam.co.jp

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HepaRG)
Specification
HepaRG
Fixative
Formaldehyde
Permeabilization
Yes - Triton X-100 in PBS
Blocking step
Milk as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: RT°C

Mrs. Alina Macovei

Verified customer

Submitted May 03 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HepaRG)
Loading amount
10 µg
Specification
HepaRG
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C

Mrs. Alina Macovei

Verified customer

Submitted May 03 2012

1-10 of 23 Abreviews or Q&A

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