製品の概要

  • 製品名
    Anti-Cathepsin B antibody [CA10]
    Cathepsin B 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [CA10] to Cathepsin B
  • 由来種
    Mouse
  • 特異性
    This antibody recognizes an epitope on the heavy chain of mature Cathepsin B and therefore detects both mature (~31 kDa) and pro (~43 kDa)Cathepsin B.
  • アプリケーション
    適用あり: IHC-P, Flow Cyt, WB, IP, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Cow, Human, Pig
  • 免疫原

    Cathepsin B purified from human liver

  • エピトープ
    Within the heavy chain of mature cathepsin B
  • ポジティブ・コントロール
    • HepG2 cells

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab58802 in the following tested applications.

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

アプリケーション Abreviews 特記事項
IHC-P 1/200.
Flow Cyt Use 0.5µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

WB Use a concentration of 0.25 µg/ml. Predicted molecular weight: 38 kDa.
IP Use a concentration of 1 µg/ml.
ICC/IF Use at an assay dependent concentration.

ターゲット情報

  • 機能
    Thiol protease which is believed to participate in intracellular degradation and turnover of proteins. Has also been implicated in tumor invasion and metastasis.
  • 配列類似性
    Belongs to the peptidase C1 family.
  • 細胞内局在
    Lysosome. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Amyloid precursor protein secretase antibody
    • APP secretase antibody
    • APPS antibody
    • CATB_HUMAN antibody
    • Cathepsin B heavy chain antibody
    • Cathepsin B1 antibody
    • CathepsinB antibody
    • CPSB antibody
    • CTSB antibody
    • cysteine protease antibody
    • OTTHUMP00000116009 antibody
    • OTTHUMP00000229510 antibody
    • OTTHUMP00000229511 antibody
    • OTTHUMP00000229512 antibody
    • OTTHUMP00000229514 antibody
    • OTTHUMP00000229515 antibody
    • OTTHUMP00000229516 antibody
    • Preprocathepsin B antibody
    see all

画像

  • Immunofluorescence analysis of U87MG cells incubated with netrin-1, staining Cathepsin B with ab58802.
  • Overlay histogram showing HepG2 cells stained with ab58802 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab58802, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • ab58802 staining Cathepsin B in murine mammary gland tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    Tissue was fixed in formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer. Samples were then blocked and then incubated with ab58802 at a 1/200 dilution for 30 minutes at 22°C. The secondary used was a biotin conjugated rabbit anti-mouse monoclonal at a 1/250 dilution.

    See Abreview

参考文献

This product has been referenced in:
  • Hsu SPC  et al. Temozolomide, sirolimus and chloroquine is a new therapeutic combination that synergizes to disrupt lysosomal function and cholesterol homeostasis in GBM cells. Oncotarget 9:6883-6896 (2018). Read more (PubMed: 29467937) »
  • Tang Y  et al. Cathepsin B inhibition ameliorates the non-alcoholic steatohepatitis through suppressing caspase-1 activation. J Physiol Biochem N/A:N/A (2018). Read more (PubMed: 30019185) »
See all 33 Publications for this product

レビューと Q&A

1-10 of 15 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (gall bladder cancer cell line OCUG-1)
Gel Running Conditions
Reduced Denaturing (12% SDS PAGE)
Loading amount
50 µg
Specification
gall bladder cancer cell line OCUG-1
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C

Mr. Siddharth Mehra

Verified customer

投稿 Jun 01 2016

Answer

The immunogen used to create ab58802 is purified cathepsin B from human liver. The epitope has been mapped to the heavy chain of mature cathepsin B, but no more specifics are known.

We have no data showing whether or not this antibody will cross react with the other cathepsins. That being said, the monoclonal antibody has a very long and trusted publication history:
http://scholar.google.com/scholar?hl=en&q=ca10+cathepsin&btnG=&as_sdt=1%2C22&as_sdtp=


If you would like to try this antibody in rabbit, you could participate in our AbTrial program to test it risk free. Please contact me if you're interested: www.abcam.com/abtrial

Read More

Answer

I am sorry to hear that you have been experiencing problems using this product in the application that you wish.

In order to assess the quality of our products I would ask that you complete a brief questionnaire relating to the application used. Often it is possible to make suggestions that may help resolve problems experienced using a particular product.

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at fault the technical support team will consider removing the product from our catalogue in order to avoid future customer inconvenience.

Could you provide some further details of the protocol used and complete the following form (attached as a word document). It would be much appreciated if you could attach an image to the response.

Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

Read More

Answer

Thank you for contacting us.

Unfortunately, alignment of our immunogen with mosquito Cathepsin B shows identity below threshold for reactivity of this antibody in this species. An immunogen should have 85% identity with the target of interest. I was able to repeat this alignment with each of our Cathepsin B antibodies and did not find one which we could recommend for use with mosquito. As such these products would not be eligible for our AbTrial program.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

I am very pleased to hear you would like to test ab5076, ab8049, ab18258, ab58802, ab64581, ab82259 and send us your results. The testing discount doesn't apply to ab4674 since this has already been tested in Rhesus monkey and IHC so we would guarantee that experiment to work.

The codes will give you a value off your next orders before the expiration date. To redeem this offer, please submit 6 Abreview with rhesus monkey data and include the code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

The code will be active once the Abreview wit has been submitted and can be redeemed in one of the following ways:
1) Call to place your order and mention the code to our customer service department;
2) Include the code in your fax order;
3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated. Remember if the product does not work you will be covered by our Abpromise guarantee and eligible for a full refund or replacement.

If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/abtrial.

Read More

Answer

Thank you for taking time toanswer my questions.

I would like to offer some suggestions to help optimize the results:

1. First thing first: ab77355 is a goat antibody. The secondary antibody will cross reacts with Bovine Ig present in BSA due to a high homology. Therefore, use the serum of the secondary as a blocking agentif possible. http://www.immunoportal.com/modules.php?name=gallery2&g2_itemId=9222 This should help a lot to lower the background.

2. In order to avoid over-loading and therefore to decrease the side bands, I would like to suggest loading 20 to 30 ug protein per lane.

3. As the blocking agent is a protein itself, prolonged blocking and in a high concentration may lead to some unwanted adherence to the blot. A maximum of 2h with 5% BSA should be enough and avoid this phenomena.

4. To clear up the image it is advisable to filter the blocking solution prior to incubation.

5. Please decrease the concentration of ab77355 down to 1:5000 to increase the specificity and to lower the background.

6. Please be aware that both antibodies needs to be stored at -20C. ab58802: Lyophilized antibodies should be resuspended at 4°C with occasional gentle mixing for at least 2 h. Store at 4°C until reconstituted, then store in aliquots at -20°C or at 4°C with 0.1% azide.

7. Please perform an incubation with the secondary antibody only to check if the secondary reacts with the sample.


Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

Read More

Answer

Thank you for contacting us.

Your credit note ID is ******.

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

Read More

Answer

Thank you for taking time to check these additional details. The details provided will enable us to investigate this case. Itam very sorrythatthe results with this antibodyhave not been successful.

I have checked the information provided and compared the two antibodies. Indeed, as the ab58802 is a monoclonal antibody, it should provide not more bands than the polyclonal antibody. It appears thereforethat you may have received a faulty vial.

I apologize for the inconvenience and am pleased to offer you a free of charge replacement or acredit note.

Thank you for your cooperation. I look forward to hearing from you with details of how you and the customerwould like to proceed.

Read More

Question

xxAbcam product code: ab58802 - Antibody storage conditions (temperature/reconstitution etc) -20℃ 2. Please describe the problem (high background, wrong band size, more bands, no band etc). More bands 3. On what material are you testing the antibody in WB? - Species: Human - What’s cell line or tissue:293T - Cell extract or Nuclear extract: cell extract - Purified protein or Recombinant protein: Overexpress recombinant protein 3. The lysate - How much protein was loaded: 20 ug - What lysis buffer was used: - What protease inhibitors were used: - What loading buffer was used: - Phosphatase inhibitors - Did you heat the samples: temperature and time: 4. Electrophoresis/Gel conditions/ Transfer conditions - Reducing or non reducing gel: non - Reducing agent: - Gel percentage : 10% SDS-PAGE - Transfer conditions: (Type of membrane, Protein transfer verified): NC paper 5. Blocking conditions - Buffer: TBST - Blocking agent: milk, BSA, serum, what percentage: 5% milk - Incubation time: 1 hr - Incubation temperature: RT 6. Primary Antibody - Species:mouse - Reacts against: human - At what dilution(s) have you tested this antibody:1:400 - What dilution buffer was used: TBST - Incubation time: O/N - Incubation temperature: 4℃ - What washing steps were done: 7. Secondary Antibody - Species: - Reacts against: mouse - At what dilution(s) have you tested this antibody: 1:10000 - Incubation time: 1 hr - Wash steps: 15 min×3 times - Fluorochrome or enzyme conjugate: - Do you know whether the problems you are experiencing come from the secondary? 8. Detection method ECl, ECl+, other detection method: 9. Did you apply positive and negative controls along with the samples? Please specify. 10. Optimization attempts - How many times have you tried the Western? 3 times - Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): no - Do you obtain the same results every time e.g. are background bands always in the same place? yes - What steps have you altered? Concentration of primary antibody

Read More
Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like appreciate if you can confirm some further details:

1.) Have the same lysates been used to compare the two antibodies?

2.) Indeed, the difference of bands is obvious between the two antibodies used. In order to know whether oneantibody can recongize morepotential forms, can you pleaseprovide me with the datasheet of the other antibody so I cancompare whether the epitopes of the two antibodies are in the same region?

3.) Has the customer also used reduced samples?

As you know, in the case that we can not find a solution to the problem and if we have to conlcude that the antibody is not functioning as described on the datasheet, we will be pleased to provide a replacement.

Thank you verymuch for your cooperation! I am looking forward toyour reply.

Read More

1-10 of 15 Abreviews or Q&A

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

登録