Overview

  • Product name
    Anti-Serum Amyloid P / SAP antibody
    See all Serum Amyloid P / SAP primary antibodies
  • Description
    Rabbit polyclonal to Serum Amyloid P / SAP
  • Host species
    Rabbit
  • Specificity
    Specific for human Serum Amyloid P / SAP with no crossreactivity with CRP or other pentraxins.
  • Tested applications
    Suitable for: WB, ELISA, Sandwich ELISAmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Full length native protein (purified) corresponding to Human Serum Amyloid P/ SAP.

  • General notes

    Previously labelled as Serum Amyloid P.

Properties

Applications

Our Abpromise guarantee covers the use of ab17872 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 1/250 - 1/2500. Predicted molecular weight: 26.76 kDa.
ELISA 1/2500 - 1/10000.
Sandwich ELISA Use at an assay dependent dilution. Can be paired for Sandwich ELISA with Mouse monoclonal [14B4] to Serum Amyloid P / SAP (ab27313). Use as detection antibody with recommended pair. See Abreview submitted by Jennifer Webster for details.

Target

  • Function
    Can interact with DNA and histones and may scavenge nuclear material released from damaged circulating cells. May also function as a calcium-dependent lectin.
  • Tissue specificity
    Found in serum and urine.
  • Involvement in disease
    Note=SAP is a precursor of amyloid component P which is found in basement membrane and associated with amyloid deposits.
  • Sequence similarities
    Belongs to the pentaxin family.
    Contains 1 pentaxin domain.
  • Post-translational
    modifications
    N-glycosylated with a complex biantennary oligosaccharide chain with a sialic acid at the end (disialo-SAP). Monosialo-SAP as well as asioalo-SAP are also detected (PubMed:15174148).
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • 9.5S alpha 1 glycoprotein antibody
    • 9.5S alpha-1-glycoprotein antibody
    • Amyloid P component serum antibody
    • APCS antibody
    • MGC88159 antibody
    • Pentaxin related antibody
    • Pentraxin 2 antibody
    • PTX 2 antibody
    • PTX2 antibody
    • SAMP_HUMAN antibody
    • SAP antibody
    • Serum Amyloid P antibody
    • Serum amyloid P component antibody
    • Serum amyloid P-component(1-203) antibody
    see all

References

ab17872 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

The reason why the customer is not seeing any bands is that normal brain has very very little SAP protein, therefore it is not surprising that the customer still sees no band. The reference below should be very useful, in particular, figure 3, which shows the levels of SAP in human brain lysate compared to an AD brain lysate: Human neurons generate C-reactive protein and amyloid P: upregulation in Alzheimer’s disease. Yasojima et al. Brain Research 887 (2000) 80–89. I therefore believe that the problem is not due to the antibody but to the low levels of protein in the customer's samples. I cannot offer a refund/credit note for this reason, Thank you for explaining this to your customer. Should she/he experience problems with a samples containing high levels of protein please do not hesitate to contact me again,

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Answer

As Serum Amyloid P protein is only expressed in high levels in AD patients as is in low levels in normal subjects I think the problem is therefore low levels in the samples. Furthermore, the customer should try incubating in more concentrated antibody (still overnight) as the recommended range is 1/250-1/2500. I think it is essential for the customer to run a positive control to determine how high the levels of serum amyloid P protein are in their samples. Please do not hesitate to contact me for further assistance,

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Question

Our customer had some trouble of ab17872 in western. They check the secondary antibody already by other primary antibodies, and the data showed that the secondary antibody could work well. Because of the film showed that ab17872 could interact with positive control in the third time., they thought the antigen binding ability of this antibody is OK. Maybe this is due to the specificity of this antibody. We would be appreciated if you could help them to deal with this situation. Thank you! 1. Order details: a.. Batch number (lot No.): 175523 b.. Abcam order or Purchase order number: ab17872 c.. Antibody storage conditions: store at -20? 2. Please describe the problem (high background, wrong band size, more bands, no band etc). We followed the Abcam WB protocol (download from the website www.abcam.com/technical) to do our experiment; we found the WB result had some problem: wrong band size and more bands in the 3 times experiments, we also found the high back ground in the second time experiment. 3. On what material are you testing the antibody in WB? · Species: Homo Sapiens · Cell extract or Nuclear extract: human plasma. · Purified protein or Recombinant protein: purified protein. 3. The lysate a.. How much protein was loaded: 30 mg a.. What lysis buffer was used: no b.. What protease inhibitors were used: no c.. What loading buffer was used: we used the sample buffer include 2%SDS, 125 mM Tris, EDTA. 2Na 2mM and 5% beta-mercaptoethanol. d.. Did you heat the samples: temperature and time: yes, we heat the samples at 95? in five minutes. 4. Electrophoresis/Gel conditions/ Transfer conditions a.. Reducing or non reducing gel: reducing gel b.. Gel percentage : 12.5% SDS-PAGE c.. Transfer conditions: according to the gel size for mA calculation and one hour for transferred. 5. Blocking conditions a.. Buffer: PBS b.. Blocking agent: milk, BSA, serum, what percentage: 5% BSA for blocking. c.. Incubation time: O/N d.. Incubation temperature: 4? 6. Primary Antibody a.. Specification (in which species was it raised against): we used the rabbit polyclonal to serum Amyloid P (ab17872) · At what dilution(s) have you tested this antibody: 1/2500X · What dilution buffer was used: 5% BSA blocking buffer. · Incubation time: one hour · Incubation temperature: RT · What washing steps were done: 0.05% PBST washed three times, each time 10 minutes. 7. Secondary Antibody a.. Specification (in which species was it raised against)? We used the goat anti rabbit IgG-HRP as secondary antibody. b.. At what dilution(s) have you tested this antibody: 1/10000X c.. Incubation time: one hour. d.. Wash steps: 0.05% PBST washed four times, each time 10 minutes. e.. Do you know whether the problems you are experiencing come from the secondary? I think this problem maybe not happened in our experiment. 8. Detection method ECl, ECl+, other detection method: ECL for developed. 9. Background bands · Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): we also used the Abcam ab27313 for the experiment, the same secondary antibody we used, but we only saw the high background in this polyclonal antibody. · Is the blocking step sufficient? Yes · Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) Yes · At what size are the bands migrating? Could they be degradation products of your target? 50 kDa, 130 kDa and above 170 kDa. · Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient) please see the attached file. 11. Did you apply positive and negative controls along with the samples? Please specify. Yes, we use the human serum purified serum Amyloid P as positive control in the experiments. (Please check the attached file.) 10. Optimization attempts · How many times have you tried the Western? Three times. · Do you obtain the same results every time e.g. are background bands always in the same place? Yes. (Please check the attached please.) · What steps have you altered? All the time, we followed the Abcam protocol.

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Answer

Thank you for contacting us for technical support on behalf of your customer. Many thanks for providing so much protocol information and the images of the blot. It enabled me to understand the problem very easily and rapidly. I am a little confused that the researcher seems different signals in the three experiments, are they the same samples, or different samples? I think the researcher's protocol is very good, I would expect the band size to be slightly lower than that seen by the monoclonal antibody that the researcher use. Is the researcher therefore confident that the higher band size detected by the monoclonal is the correct one? I am guessing that the serum samples are from AD patients, not control patients? I think the main problem is that the membrane is unfortunately blocked too much and the antibody added only for a short amount of time but in a very concentrated manner. I would therefore recommend, like in our lab, to incubate the membrane for 1 hour (it may be worth trying milk too as well as 5% BSA) and incubating the membrane overnight, probably in more diluted antibody (in TBST only). This will promote a slow but targeted binding of the antibody to the Serum Amyloid P protein. If the customer still experiences problems please do not hesitate to contact me again and I can offer a replacement vial or refund if the antibody was purchased in the last 90 days,

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ELISA
Sample
Human Serum (Serum)
Specification
Serum
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Type
Sandwich

Miss. Jennifer Webster

Verified customer

Submitted Aug 16 2006

Question
Answer

Good morning, Unfortunately the source of ab17872 does not have an image of the western blot to provide us. I am very sorry for this inconvenience, Best of luck with your research,

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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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