Overview

  • Product name
  • Description
    Rabbit polyclonal to UBPY/USP8
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ELISA, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (Human) conjugated to KLH selected within aa 1050~1100 of human USP8.

  • Positive control
    • Human breast carcinoma and hepatocarcinoma tissues and transfected 293T cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab38865 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
ELISA 1/1000.
WB 1/100 - 1/500. Detects a band of approximately 128 kDa (predicted molecular weight: 128 kDa).
IHC-P 1/50 - 1/100.

Target

  • Function
    Hydrolase that can remove conjugated ubiquitin from proteins and therefore plays an important regulatory role at the level of protein turnover by preventing degradation. Converts both 'Lys-48' an 'Lys-63'-linked ubiquitin chains. Catalytic activity is enhanced in the M phase. Involved in cell proliferation. Required to enter into S phase in response to serum stimulation. May regulate T-cell anergy mediated by RNF128 via the formation of a complex containing RNF128 and OTUB1. Probably regulates the stability of STAM2 and RASGRF1. Regulates endosomal ubiquitin dynamics, cargo sorting, membrane traffic at early endosomes, and maintenance of ESCRT-0 stability. The level of protein ubiquitination on endosomes is essential for maintaining the morphology of the organelle. Deubiquitinates EPS15 and controles tyrosine kinase stability. Removes conjugated ubiquitin from EGFR thus regulating EGFR degradation and downstream MAPK signaling. Involved in acrosome biogenesis through interaction with the spermatid ESCRT-0 complex and microtubules. Deubiquitinates BIRC6/bruce and KIF23/MKLP1.
  • Sequence similarities
    Belongs to the peptidase C19 family.
    Contains 1 MIT domain.
    Contains 1 rhodanese domain.
    Contains 1 USP domain.
  • Domain
    The MIT domain is required for endosomal localization, CHMP1B-binding, maintenance of ESCRT-0 stability and EGFR degradation.
    The rhodanese domain is sufficient for RNF41-binding.
  • Post-translational
    modifications
    Phosphorylation of Ser-718 is essential for interaction with YWHAE and for cytosol localization. Undergoes dephosphorylation at Ser-718 in the M phase. Tyrosine-phosphorylated in its N-terminal half in an EGFR-dependent manner.
    Ubiquitinated. Inactive form is mostly monoubiquitinated, but polyubiquitination happens too. Ubiquitination is increased in EGF-stimulated cells. Ubiquitination of active form is undetectable, suggesting a possibility that USP8 deubiquitinates itself, thereby regulating its own function.
  • Cellular localization
    Cytoplasm. Nucleus. Endosome membrane. Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Deubiquitinating enzyme 8 antibody
    • FLJ34456 antibody
    • hUBPy antibody
    • HumORF8 antibody
    • KIAA0055 antibody
    • MGC129718 antibody
    • SPG59 antibody
    • Ubiquitin carboxyl-terminal hydrolase 8 antibody
    • Ubiquitin isopeptidase Y antibody
    • ubiquitin specific peptidase 8 antibody
    • Ubiquitin thioesterase 8 antibody
    • Ubiquitin-specific-processing protease 8 antibody
    • UBP8_HUMAN antibody
    • UBPY antibody
    • Usp8 antibody
    see all

Images

  • Anti-UBPY/USP8 antibody (ab38865) + WiDr cell lysate

    Predicted band size: 128 kDa

  • Formalin fixed and paraffin embedded human breast carcinoma tissue stained with ab38865, followed by AEC staining.

References

ab38865 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Question

Dear Kate   Further to our telephone conversation please find below the responses to the questions you emailed me last week and two images of the WB with anti-UBPY antibody.   As I mentioned on the phone, I have purchased many antibodies from abcam and I am always very satisfied with them. However, based on our extensive experience with immunoblotting, I feel that this antibody is not working and I would appreciate it if we could be reimbursed or given teh option to buy another product.   I look foward to hearing from you best wishes       Order Details Antibody code: ab28865 Problem: No distinct band seen on western blot as advertised on your website (see attached images) Choose: Multiple bands  Lot number: 866221 Purchase order number: 12703 (may not be the complete number) ordered on the 05-09-11 General Information Antibody storage conditions (temperature/reconstitution etc) The antibody was aliquoted and frozed on arrival. Aliquots were thawed and used once or twice. Description of the problem (high background, wrong band size, more bands, no band etc.)   Multiple bands. As you can see from the WB no distinct band is seen at the correct molecular weight. In fact in brain strong bands are seen at the wrong MW Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.) HEK cell extract and human brain lysate Sample preparation (Buffer/Protease inhibitors/Heating sample etc.) Cells or brain tissue were homogenised in ice-cold 100mM KCL, 10mM Tris pH7.4 containing 1% Triton X, 0.1% SDS, complete protease inhibitor cocktail, 1mM N-ethyl-maleimide and 1mM PMSF. Lysates were left on rotator for 20min at 4 C and then centrifuged at 13000rpm for 5min at 4 C. The supernatant was quantified and 50-100 micrograms of protein were mixed with LDS and heated at 95 C for 6min before loading on Invitrogen minigels.  Amount of protein loaded 50-100micrograms Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.) 4-12% Bis-Tris NuPAGE (Invitrogen) Transfer and blocking conditions (Buffer/time period, Blocking agent etc.) Running and transfer buffers were purchased from Invitrogen and are routinely used in our laboratory block for 1h in 4% milk Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) ab38865 range of dilutions used. 1:100, 1:200, 1:500 Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) ECL peroxidase conjugated anti-rabbit Detection method (ECL, ECLPlus etc.) ECL (GE) Positive and negative controls used (please specify) Negative control: no primary Positive control: UBPY/USP8 should be detected in human brain Optimization attempts (problem solving) used different blocking conditions and primary antibody dilutions How many times have you tried the Western? 5 times Have you run a "No Primary" control? Yes Do you obtain the same results every time? e.g. are the background bands always in the same place? Yes What steps have you altered? as above Additional Notes Image: see attachement        

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Answer

Thank you for your message and for providing this further information. Reviewing the details, there are regrettably no major suggestions to provide to help improve the results on this occasion. I appreciate the time you have spent on these experiments and would be pleased to arrange a refund or credit note or free of charge replacement in compensation.  For free of charge replacements, we are happy to provide alternatives. The only minor points I can recommend to consider for future experiments are as follows: 1. We recommend to load 20 - 30 ug of protein per lane of the gel, to ensure the gel is not overloaded which can give background staining. 2. I can suggest to try incubation overnight at 4oC, also to try 1:1000 lower concentration, this should help provide more specific staining. 3. I can recommend to consider if the secondary antibody working well with other antibodies? I look forward to hearing from you with details of how you would like to proceed.  

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Question
Answer

Thank you for your telephone call this morning. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund. As discussed on the telephone, I would like to investigate this further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. I would appreciate if you could also provide an image which would help us to assess the results. Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.   Order Details Antibody code: Problem Choose: Non-specific band Multiple bands No signal or weak signal High background Lot number Purchase order number or preferably Abcam order number: General Information Antibody storage conditions (temperature/reconstitution etc) Description of the problem (high background, wrong band size, more bands, no band etc.) Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.) Sample preparation (Buffer/Protease inhibitors/Heating sample etc.) Amount of protein loaded Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.) Transfer and blocking conditions (Buffer/time period, Blocking agent etc.) Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Detection method (ECL, ECLPlus etc.) Positive and negative controls used (please specify) Optimization attempts (problem solving) How many times have you tried the Western? Have you run a "No Primary" control? Yes No Do you obtain the same results every time? Yes No e.g. are the background bands always in the same place? What steps have you altered? Additional Notes: Image: We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asses the results.  

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