Overview

  • Product name
    Anti-S100 antibody [8B10]
    See all S100 primary antibodies
  • Description
    Mouse monoclonal [8B10] to S100
  • Host species
    Mouse
  • Specificity
    Specific to S-100 beta beta and alpha beta. It does not recognize the alpha-alpha form or alpha subunit only.
  • Tested applications
    Suitable for: ELISA, IHC - Wholemount, ICC/IF, WB, Sandwich ELISAmore details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Full length protein (Human).

  • Positive control
    • Human brain.
  • General notes

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab868 as a replacement.

Properties

Applications

Our Abpromise guarantee covers the use of ab8330 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 Use at an assay dependent concentration.
IHC - Wholemount Use at an assay dependent concentration. PubMed: 21856924
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 21 kDa.
Sandwich ELISA Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Mouse monoclonal [6G1] to S100 (HRP) (ab10203). Sensitive to EDTA or other bivalent-binding agents. Better performance can be obtained in presence of 5mM CaCL2 in these assay buffers. Can be used as capture antibody in conjunction with ab10203.

Target

  • Function
    Weakly binds calcium but binds zinc very tightly-distinct binding sites with different affinities exist for both ions on each monomer. Physiological concentrations of potassium ion antagonize the binding of both divalent cations, especially affecting high-affinity calcium-binding sites.
  • Tissue specificity
    Highly prevalent in heart. Also found in lesser quantities in skeletal muscle and brain.
  • Sequence similarities
    Belongs to the S-100 family.
    Contains 2 EF-hand domains.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • Bpb antibody
    • NEF antibody
    • Protein S100-A1 antibody
    • S-100 protein alpha chain antibody
    • S-100 protein subunit alpha antibody
    • S100 alpha antibody
    • S100 beta antibody
    • S100 calcium binding protein A1 antibody
    • S100 calcium binding protein B antibody
    • S100 calcium-binding protein A1 antibody
    • S100 protein alpha polypeptide antibody
    • S100A antibody
    • s100a1 antibody
    • S100B antibody
    • S100beta antibody
    • S10A1_HUMAN antibody
    see all

Images

  • Interaction of several monoclonal antibodies with purified S100 protein (1 µg) from human brain in WB, after native gel electrophoresis using the Ornstein-Davis system. Lane 1 = ab14849, Lane 2 = ab8330, ab Lane 3 = ab10203, Lane 4 = ab8334. The doublet band appears at about 20-21 kD, Interaction of several monoclonal antibodies with purified S100 protein (1 µg) from human brain in WB, after native gel electrophoresis using the Ornstein-Davis system. Lane 1 = ab14849, Lane 2 = ab8330, ab Lane 3 = ab10203, Lane 4 = ab8334. The doublet band appears at about 20-21 kD,

  • S-100 calibration curve, from one step assay in streptavidin coated plates. Ab8330 [200 ng/well] was used as capture and Eu- labelled ab10203 [200 ng/well] as detection. Antigen used was S-100 protein from the human brain. Incubated 20 minutes at 20°C.

References

This product has been referenced in:
  • Wu H  et al. N-Terminal Domain of Feline Calicivirus (FCV) Proteinase-Polymerase Contributes to the Inhibition of Host Cell Transcription. Viruses 8:N/A (2016). Read more (PubMed: 27447663) »
  • deBlacam C  et al. HOXC11-SRC-1 regulation of S100beta in cutaneous melanoma: new targets for the kinase inhibitor dasatinib. Br J Cancer 105:118-23 (2011). IHC . Read more (PubMed: 21654685) »
See all 6 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Sample
Mouse Tissue sections (Embryonic and postnatal mouse cerebellum)
Specification
Embryonic and postnatal mouse cerebellum
Permeabilization
Yes - 0.5% Triton in PBS
Fixative
Paraformaldehyde

Qiuxia Guo

Verified customer

Submitted Nov 01 2013

Answer

Thank you for contacting us and sorry for the delay in getting back to you.

Kate is unfortunately still not in the office but I will try to answer your questions as best I can.

I can confirm that the antibodies mentioned (ab8330 and ab10203; ab8334 and ab10203) have been shown to work as a pair in sandwich ELISA.

In answer to your following questions, it is regrettable that we are not able to provide the standard ab95190. I am sorry but I am not able to suggest an alternative standard from a different supplier. In answer to your question regarding the forms of the protein the antibodies is likely to recognise, unfortunately, only a limited amount of epitope mapping has been performed and the information which has been gained from these experiments has been added to the datasheets of the antibodies.

As you have seen, the ab8330 has been shown to be able to recognise the beta-beta and the alpha-beta form but is not able to recognise the alpha-alpha or the alpha subunit only. Whilst ab8334 has been shown to be specific for the alpha-beta complex and the ab10203 is able to recognise the beta-beta and alpha-beta complex. Further than this we have not investigated the binding properties of the antibodies.

I hope this information has been of some help. If I can be of any further assistance, please do not hesitate to let me know.

Read More

Answer

Thank you for your reply.

My sincere apologies for the confusion. In this case, I am sorry we do not have a suitable standard protein to suggest on this occasion.

In answer to your second question regarding ab48653 and ab105371, I am sorry these have not been tested in sandwich ELISA and regrettably would not be covered by the guarantee for this application. They have been tested only in indirect ELSA for each antibody against its antigen.

I am sorry to confirm that because we carry over 90 000 products, it is not possible for use tostock sample vials to send to customers. We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase. If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Please contact me again by replying to this email prior to purchase if you would like more information.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contactme again with any other questions

Read More

Answer

Thank you for your follow up message, and I appreciate your patience waiting for a response. I have been collecting information from the originator of these antibodies.

Reviewing the details for our S100 antibodies, I can confirm the following recommended matched pairs for sandwich ELISAas stated on the datasheets:

ab8330: Tested in human. Can be paired for Sandwich ELISA with Mouse monoclonal [6G1] to S100 (HRP) (ab10203), Can be used as capture antibody in conjunction with ab10203.

ab8334: Tested in human. Can be paired for Sandwich ELISA with Mouse monoclonal [6G1] to S100 (HRP) (ab10203). Can be used as capture antibody in conjunction with ab10203.

Sandwich ELISA procedures can be difficult to optimize and so we recommend tested match pair antibodies should be used. This ensures the antibodies are detecting different epitopes on the target so they do not interfere with the other antibody binding. Please note we are unable to guarantee our antibodies in sandwich ELISA unless they have been specifically tested together in sandwich ELISA.

For a standard, we can recommend to use ab95190 - S100 protein (Human)
https://www.abcam.com/index.html?datasheet=95190 (or use the following: https://www.abcam.com/index.html?datasheet=95190).

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for your inquiry.

I am happy to confirm that we do have an antibody in our catalogue that can be used as astrocyte marker and is tested and guaranteed for IHC- whole mounts:

ab8330

https://www.abcam.com/index.html?datasheet=8330 (or use the following: https://www.abcam.com/index.html?datasheet=8330).

Please follow the above link to review the datasheet.

Please do not hesitate to contact me again with any further questions.

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 to offer some suggestions to help optimize the results from ab955 since this antibody has been tested in mouse paraffin-embedded tissue sections.


For the fixation, we generally recommend between 18 - 24 hrs. It may be just that your sections are underfixed.


For antigen retrieval, was this performed in the microwave? We generally recommend 20 mins, but it's best to optimize between 15 - 30 mins. When the 20 mins has elapsed, you can run cold tap water into the vessel for 10 mins.


If you want to permeablize the sections, you should do a 0.1% Triton-X 100 in PBS treatment as a separate step and omit the Triton X-100 from the blocking buffer.


For blocking, have you tried5-10% normal goat serum instead of BSA? You should also put this in PBS.


The most important thing to try wouldbe to test a1/100 dilution of ab955 overnight at 4C instead of 1/200 for 1 hr RT. You should do this incubation in 1% BSA in PBS.


For the s100 antibody ab8330, would you be able to run a positive control with rat tissues since this antibody isn't tested in mouse?


For the CD31 antibody, ab7388, we know that this does not work well with PFA fixation (see Abreview linkedbelow). We have heard that it works wellwhen used on frozen sections of mouse skin that werefixed in methanol:acetone for 10 mins. Would you be able to try some frozen methanol or acetone fixed sections as a positive control since IHC-P does not work.


https://www.abcam.com/index.html?datasheet=7388&tab=abreviews&intabreviewid=23226

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

In the event that a ab955 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 for this tested antibody.

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

Read More

Answer

I have contacted the lab regarding the epitope for ab8330 and received the following answer. "Unfortunately we have not determined the epitope for anti-S100 ab8330. The only information we have is that according to our internal tests ab8330 recognizes alpha-beta and beta-beta forms. It does not recognize the alpha-alpha form or alpha subunit only." I am sorry, that I could not give you more detailed information and look forward to receiving your questionnaire.

Read More

Answer

Thank you for your enquiry and also your patience. I would actually recommend that you pair ab8330 and ab10203 or ab8334 and ab10203. I will update the datasheets to reflect this also. I have also included our protocol for use with these antibodies. Either pair of antibodies should work for you and I would be interested in hearing your feedback if you purchase these antibodies. Here is our protocol: Two-step “sandwich” immunofluorometric assay for S-100 protein: Assay should be performed in 96-well microtitration plates. Capture Mabs are unlabeled, detection Mabs are labeled with the stable chelate of Eu3+. 1st step Capture Mabs (1 µg/well; 100 µl/well) are incubated in PBS (10 mM KH2PO4, 150 mM NACL, pH 7.4) in 96-well plate for 60 minutes at room temperature with gentle shaking. 2nd step After washing (DELFIA® washing solution, two times) the mixture of antigen (30 µl/well, standards, dissolved in normal human serum or testing samples) and detection Mabs (100 µl/well, in DELFIA® Assay Buffer) are added. After 30 minutes incubation at room temperature the plates are washed 6 times, LANFIA enhancement solution (0.2 ml/well) is added, and incubation is continued for 3 minutes at room temperature with gentle shaking. The fluorescence (cps) is measured with the 1234 DELFIA® Plate Fluorometer. The method described has a detection limit below 0.05 µg/L, and the linearity range is 0.1–500 µg/L. I hope this information helps, please do not hesitate to contact us if you need any more advice or information,

Read More

Answer

Thank you for your reply. We would like to confirm that ab8330 recognizes both the alpha-beta and beta-beta forms of S-100 in Western blotting. A western blotting image with the legend can be seen on our on-line product datasheet. Western blotting protocol - protein samples are separated by 10-20 % gradient SDS gel electrophoresis and transferred to nitrocellulose membrane (Biotechnology, 1992, 24: 145-9). To block nonspecific binding of the proteins, the nitrocellulose membrane was incubated for 15 min at room temperature in phosphate buffered saline, containing 1 g/L Tween 20 and 5 g/L casein. The membrane was then incubated for 1 hour in the same buffer containing 0.02 g/L anti- S100 antibodies and 1 g/L casein. After the nitrocellulose membrane was washed with phosphate buffered saline containing 1 g/L Tween 20, it was incubated for 2 hours with horseradish peroxidase-labelled anti-mouse antibodies, and the binding of Mabs was visualized by incubation with diaminobenzidine. We hope this information will be useful for you.

Read More

Answer

Thank you for your enquiry and your interest in our products. We agree with you, it is always very important to use positive control. Human brain lysate itself or lysate from astrocytes, Schwann's cells, ependymomas and astrogliomas can be used as positive control for this antibody. If you need anything further or any help then please let us know.

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1-10 of 11 Abreviews or Q&A

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

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