Overview

  • Product name
    Anti-Cathepsin D antibody, prediluted
    See all Cathepsin D primary antibodies
  • Description
    Rabbit polyclonal to Cathepsin D, prediluted
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Cathepsin D isolated from human liver.

Properties

Applications

Our Abpromise guarantee covers the use of ab925 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
IHC-P
IHC-Fr
  • Application notes
    This antibody has been pretitered and quality controlled to work on formalin-fixed paraffin-embedded as well as acetone fixed cryostat tissue sections.
    No further titration is required.
    We suggest an incubation period of 30-60 minutes at room temperature.
    However, depending upon the fixation conditions and the staining system employed, optimal incubation should be determined by the user.
  • 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

    References

    This product has been referenced in:
    • Zheng X  et al. Role of the proteolytic hierarchy between cathepsin L, cathepsin D and caspase-3 in regulation of cellular susceptibility to apoptosis and autophagy. Biochim Biophys Acta 1783:2294-300 (2008). Read more (PubMed: 18775751) »
    See 1 Publication for this product

    Customer reviews and Q&As

    1-3 of 3 Abreviews or Q&A

    Question

    Thanks a lot for your reply. Please, see my answers below: lot number is 83806 1. Please describe the problem (high background, no staining etc). High background: when I blocked 10 min in each of step (H2O2, Avidin/Biotin and Protein Block Serum Free). OR: I did not get any staining or some times only macrophages: when I blocked by 15-20 minutes in each of step (H2O2, Avidin/Biotin and Protein Block Serum Free). 2. On what material are you testing the antibody in IHC? *Species? *Cell line? *Tissue? I tried this Ab on the Normal Human Breast and Human Breast Carcinoma. 3. How did you fix the samples? 10% NBF. *Ethanol, methanol *Acetone *Paraformaldehyde *Other 4. Did you apply antigen retrieval step? Yes, I tried two ways: put slides in jar with Citrate Buffer, ph 6.0; then I put jar in water bath for: a) 20 min at 98°C + 20 min to cooling at RT; b) 37 min at 97°C + 12 min to cooling at RT. *Enzymatic method *Heat mediated technique *Other 5. How did you block the unspecific binding sites? Please, see answers #1 6. Primary antibody We would like to have stain of fibroblasts and the myoepithelial cells of breast. *Specification (in which species was it raised against)? *At what dilution(s) have you tested this antibody? This Ab ready to use (ab925-6) *Incubation time is 60 min as suggested, wash steps is 2x with Tris Buffer (using autostaining machine from Richard-Allan company) (multiple short washes are more effective than fewer longer wash steps)? 7. Secondary antibody *What secondary antibody are you using? I have tried two different: Anti-mouse/rabbit from Richard Allan as well as Anti-Rabbit from Vector in different concentration from 2-10 µg/ml for 20-30 min. *Specification (in which species was it raised against)? *At what dilution(s) have you tested this antibody? *Incubation, wash steps? 2x with Tris Buffer. *Do you know whether the problems you are experiencing come from the secondary? *What detection method are you using? HP/DAB 8. Background staining *Please provide an image of your staining. Sorry, I don't have these images right now. 9. Which detection system did you use? HP/DAB 10. Did you apply positive and negative controls along with the samples? Please specify. I used the Normal Human Breast and Human Breast Carcinoma for a positive control and, the Negative Rabbit IgG for a negative control in concentration of 0.10mg/ml. 11. Optimization attempts *How many times have you tried the IHC? As you can see from my explanation, not at once. *Do you obtain the same results every time? Unfortunately, yes. *What steps have you altered? Every time is different steps. I hope you will be able to figurate what is going on and help me in my staining:-)

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    Answer

    Thank you for the details that you have provided and for your patience. Are you using a home made kit to stain this antibody with? This may not be as sensitive as a commercial kit, such as Dako's or Power Vision. At this point I would suggest increasing the blocking incubation period. What were your results with the negative control?

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    Answer

    Thanks for your email. If you continue to experience difficulty with ab925 in IHC, please let me know. Just answer the questions below, and also include the lot number that you received (located on the vial), and the Abcam order number or purchase order number that was used. 1. Please describe the problem (high background, no staining etc). 2. On what material are you testing the antibody in IHC? •Species? •Cell line? •Tissue? 3. How did you fix the samples? •Ethanol, methanol •Acetone •Paraformaldehyde •Other 4. Did you apply antigen retrieval step? •Enzymatic method •Heat mediated technique •Other 5. How did you block the unspecific binding sites? 6. Primary antibody •Specification (in which species was it raised against)? •At what dilution(s) have you tested this antibody? •Incubation time, wash steps (multiple short washes are more effective than fewer longer wash steps)? 7. Secondary antibody •What secondary antibody are you using? •Specification (in which species was it raised against)? •At what dilution(s) have you tested this antibody? •Incubation, wash steps? •Do you know whether the problems you are experiencing come from the secondary? •What detection method are you using? 8. Background staining •Please provide an image of your staining 9. Which detection system did you use? 10. Did you apply positive and negative controls along with the samples? Please specify. 11. Optimization attempts •How many times have you tried the IHC? •Do you obtain the same results every time? •What steps have you altered?

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    Question
    Answer

    Thank you for your enquiry. The concentration is 0.10 mg/ml. Please note that this antibody has been pretitered and quality controlled to work on formalin-fixed paraffin-embedded as well as acetone fixed cryostat tissue sections and no further titration is required. We suggest an incubation period of 30-60 minutes at room temperature. However, depending upon the fixation conditions and the staining system employed, optimal incubation should be determined by the user. If you have any additional questions, please contact us again.

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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