Overview

  • Product name
  • Description
    Rabbit polyclonal to SPANX
  • Host species
    Rabbit
  • Specificity
    Reacts with SPANX-C, SPANX-B and SPANX-N family members.
  • Tested applications
    Suitable for: IHC-P, WB, ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Chimpanzee, Gorilla
  • Immunogen

    Full-length recombinant human SPANX-C protein.

  • Positive control
    • Nuclear extract from VWM105 cells, purified recombinant SPANX-B, SPANX-C and SPANX-N proteins.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
  • Concentration information loading...
  • Purity
    Protein A purified
  • Purification notes
    ab51493 was purified from monospecific antiserum by protein A chromatography followed by exhaustive dialysis against the buffer.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab51493 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 Use a concentration of 1.25 - 2.5 µg/ml.
WB 1/200 - 1/1000. Detects a band of approximately 19 kDa (predicted molecular weight: 11 kDa).
ELISA 1/1000 - 1/4000.
ICC/IF Use at an assay dependent concentration.

Target

  • Tissue specificity
    Detected in testis, melanoma and bladder carcinoma.
  • Sequence similarities
    Belongs to the SPAN-X family.
  • Developmental stage
    Detected in round and elongating spermatids.
  • Cellular localization
    Cytoplasm. Nucleus. Associated with nuclear craters.
  • Information by UniProt
  • Database links
  • Alternative names
    • Cancer-testis-associated protein CTp11 antibody
    • Cancer/testis antigen 11.3 antibody
    • Cancer/testis-associated protein CTp11 antibody
    • CT11.3 antibody
    • CTP11 antibody
    • Nuclear associated protein SPAN Xc antibody
    • Nuclear-associated protein SPAN-Xc antibody
    • SPANX family member C antibody
    • SPANX family, member C antibody
    • SPANX family, member C1 antibody
    • SPANX-C antibody
    • SPANXC antibody
    • Sperm protein associated with the nucleus on the X chromosome C antibody
    • SPNXC_HUMAN antibody
    see all

Images

  • All lanes : Anti-SPANX antibody (ab51493) at 1/1000 dilution

    Lane 1 : VWM105 cell nuclear extract
    Lane 2 : purified recombinant SPANX-C fusion protein
    Lane 3 : purified recombinant SPANX-B
    Lane 4 : purified recombinant SPANX-C
    Lane 5 : purified recombinant SPANX-N

    Secondary
    Lane 1 : IRDye™800 conjugated Goat anti Rabbit
    IgG [H&L]
    Lane 2 : IRDye™800 conjugated Goat anti Rabbit
    IgG [H&L]


    Predicted band size: 11 kDa
    Observed band size: 19 kDa
    why is the actual band size different from the predicted?

  • ab51493 at 2.5 ug/ml staining human sperm and spermatids at 60X. Tissue was formalin-fixed and paraffin embedded. The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain.

References

ab51493 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for your note. This is to let you know that I have just contacted and asked our Account Department to raise a credit note for you - for the cost of one vial of ab51493. For your information, the internal reference note for this credit is CNXXXXX. You can use this credit in the near future for any of the products which are in the catalogue.   I hope this helps and if I can assist further, please do not hesitate to contact me.

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Answer

Thank you for getting back to me and for passing some further results. I am very sorry to hear that even after following some tips and recommendations, the non-specific bands will not get weaker on the blot. In case your customer is dissatisfied with the quality of this product, I could certainly offer either a new vial as a free of charge replacement or a credit note. Please discuss it and let me know how you wish to proceed. I look forward to hearing from you soon.  

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Question

Dear technical support team: This customer has purchased ab51493 (Anti-SPANX antibody), and conducted the WB with human lung adenocarcinoma cell line. The results show multiple bands; therefore this customer wants to ask for your help to modify his experiment step, could you please offer any suggestion to improve his wb result? I attached the wb image in this letter and his experiment step as follow:   1. Order details: Batch number: gr45356-1 Abcam product code: ab51493 Antibody storage conditions (temperature/reconstitution etc): -20℃ 2. Please describe the problem (high background, wrong band size, more bands, no band etc).Wrong band size and more bands 3. On what material are you testing the antibody in WB? ·         Species: human ·         What’s cell line or tissue: CL 1-5(lung adenocarcinoma cell line) introduce with SPANXA1 plasmid and CL 1-0(lung adenocarcinoma cell line) only for endogenous SPANXA1 ·         Cell extract or Nuclear extract: cell extract ·         Purified protein or Recombinant protein:   3.  The lysate How much protein was loaded: 40ug What lysis buffer was used: M-PER(Thermo 78501) What protease inhibitors were used: protease inhibitor cocktail(sigma p8340) What loading buffer was used: 4x loading buffer Phosphatase inhibitors :none Did you heat the samples: temperature and time: 100℃ 10mins   4.  Electrophoresis/Gel conditions/ Transfer conditions Reducing or non reducing gel: reducing Reducing agent: SDS Gel percentage : 12% Transfer conditions: (Type of membrane, Protein transfer verified): PVDF membrane 5. Blocking conditions Buffer: TBST Blocking agent: milk, BSA, serum, what percentage:5% skim milk (BD) Incubation time: 2 hrs ·         Incubation temperature: Room temperature 6. Primary Antibody Species: rabbit Reacts against: Human SPANX ·         At what dilution(s) have you tested this antibody: 1:1000 ·         What dilution buffer was used:  5% skim milk in TBST ·         Incubation time: overnight ·         Incubation temperature: 4℃ ·         What washing steps were done: Wash with TBST three times for 10 mins   7. Secondary Antibody Species:Goat Reacts against: anti-rabbit At what dilution(s) have you tested this antibody: 1:5000 Incubation time: 30 mins in room temperature Wash steps: Wash with TBST three times for 10 mins Fluorochrome or enzyme conjugate:HRP conjugate Do you know whether the problems you are experiencing come from the secondary?  I don’t think so   8. Detection method ECl, ECl+, other detection method: FUJIFILM LAS-4000 luminascent image system 9. Did you apply positive and negative controls along with the samples? Please specify. I used CL 1-5 SPANXA1 mixed stable clone, and it can be detected by V5 antibody 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   Could you please help this customer to solve his problem? I appreciate your help. Best regards  

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Answer

Thank you for contacting Abcam Technical Team and for taking the time to provide some useful details of the experiments. I am very sorry to hear that your customer is having problems with this product. After reading through the detailed protocol you kindly forwarded to Abcam, I would like to make the following comments/suggestions: 1) It would be worth preparing nuclear lysate and run on the gel - instead of total cell lysate. 2) Please advise your customer to use positive control (purified recombinant SPANX-C). 3)  Does the detection system work fine? Have you used it successfully with another primary antibody? Have you run a no primary - only secondary antibody - control to see if any of the non-specific bands are due to the secondary or not? If you have not done yet, I would advise you to check it.   I hope this will be useful for you. Should your customer still has any problem with this antibody after following these suggestions, then please do not hesitate to contact our Technical Department again.

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Answer

Thank you for your enquiry. I have investigated this with the originator who provided this image. All the samples in lane 2 to 5 of this western blot were recombinant fusion proteins. Consequently, the molecular weights are much higher than expected for the endogenous protein. I hope this is helpful to you. Should you have any further questions, please do not hesitate to contact us.

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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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