Anti-S100 antibody [4C4.9] (ab4066)

Mouse monoclonal S100 antibody [4C4.9]. Validated in WB, IHC, ICC/IF and tested in Mouse, Rat, Cow, Human. Cited in 31 publication(s). Independently reviewed in 10 review(s).

Reviews (10)Q&A (11)References (40)

Overview

  • Product name

    Anti-S100 antibody [4C4.9]
    See all S100 primary antibodies

  • Description

    Mouse monoclonal [4C4.9] to S100

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC/IF, IHC-P, IHC-Fr, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Human

  • Immunogen

    Full length protein. Purified bovine brain S-100 protein

  • Positive control

    • Melanoma.

Properties

Applications

Our Abpromise guarantee covers the use of ab4066 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
ICC/IF Use at an assay dependent concentration. PubMed: 23624931
IHC-P 1/10 - 1/50. Antigen retrieval is not essential but may optimise staining.
IHC-Fr 1/200 - 1/500.
WB Use at an assay dependent concentration.

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

    see all

  • 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

  • Immunohistochemistry (Frozen sections) - Anti-S100 antibody [4C4.9] (ab4066)
    Immunohistochemistry (Frozen sections) - Anti-S100 antibody [4C4.9] (ab4066)This image is courtesy of an Abreview submitted by Nancy Nutile-McMenemy
    ab4066 at a 1/200 dilution staining rat spinal cord tissue sections by Immunohistochemistry (Frozen sections). Rats were transcardially perfused with 4% PFA, the tissue was post fixed 1 hour in 4% PFA then 30% sucrose for three days. 20um sections were cut with cryostat. The antibody was incubated with the tissue sections for 18 hours and then bound antibody was detected using an Alexa Fluor ® 555 conjugated goat polyclonal antibody.

    See Abreview

  • Western blot - Anti-S100 antibody [4C4.9] (ab4066)
    Western blot - Anti-S100 antibody [4C4.9] (ab4066)This image is courtesy of an Abreview submitted by Ms Nancy Nutile-McMenemy
    All lanes : Anti-S100 antibody [4C4.9] (ab4066) at 1/200 dilution

    Lanes 1-3 : 40ug Rat spinal cord homogenate
    Lanes 4-6 : 40 ug Mouse spinal cord homogenate
    Lanes 7-8 : 40 ug Rat brain homogenate

    Secondary
    All lanes : HRP conjugated goat anti-mouse

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 15,18,21 kDa
    why is the actual band size different from the predicted?


    Exposure time: 2 minutes


    The extra band at 18kDa may be a result of the protein's thermolabile properties. The band at 21 kDa maybe a dimer of s100a and s100b and the 10/15 kDa may be the subunits. However, there are also references to suggest these may be immunoreactive proteins with differing molecular weights depending on proliferative or differentiated state of the cell (Anat Embryol (Berl). 1997 Nov;196(5):403-16).

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 antibody [4C4.9] (ab4066)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S100 antibody [4C4.9] (ab4066)
    ab4066 staining human melanoma by IHC-P.

References

This product has been referenced in:

  • Lu Y  et al. Fibroblast growth factor 21 facilitates peripheral nerve regeneration through suppressing oxidative damage and autophagic cell death. J Cell Mol Med 23:497-511 (2019). Read more (PubMed: 30450828) »
  • Passipieri JA  et al. Adipose Stem Cells Enhance Nerve Regeneration and Muscle Function In A Peroneal Nerve Ablation Model. Tissue Eng Part A N/A:N/A (2019). Read more (PubMed: 30760135) »
See all 40 Publications for this product

Publishing research using ab4066? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 21 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-S100 antibody [4C4.9]

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (melanoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrite buffer, pH6.0
Permeabilization
No
Specification
melanoma
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Aug 19 2016

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-S100 antibody [4C4.9]

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin)
Antigen retrieval step
Other
Permeabilization
No
Specification
skin
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Dec 14 2015

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-S100 antibody [4C4.9]

 Inconclusive
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain (Hippocampus))
Antigen retrieval step
None
Permeabilization
Yes - 0.1%TritonX-100/PBS
Specification
Brain (Hippocampus)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.5, 2 · Temperature: RT°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted May 14 2015

Question

IHC-FoFr with rat peripheral nerve
sees no staining
fix: 4% PFA perfused, postfixed with PFA
no AR
Ab: 1/100 o/n 4 degree; 1/50 o/n RT
block: BSA + goat serum
2nd: goat anti-mouse, OK

suggested: try an enzymatic AR step

Read More
Answer


As we discussed, trying an enzymatic antigen retrieval step could help with the signal.

If not, please let me know.

As for the protocol on the datasheet - this was added most likely from a collaborator or customer. Thus, these must be the conditions that were used by them. I still would suggest to do the overnight incubation at 4 degree as this makes more sense.

Read More

Question

Dear Sir or Madam,

could you please also provide me with the concentration of your product Anti-S100 antibody [4C4.9] - Astrocyte Marker (ab4066)?

Best Regards,

Read More
Answer

Thank you for your interest in our products.

The unit size of this antibody (ab4066) is 500 µl and the concentration is not determined since it is not affinity purified - only IgG fraction.

If I can assist further, please do not hesitate to contact me.

Read More

Question

Thank you for sending the replacement antibody. This one worked very well both in vitro using human induced pluripotent stem cells and in vivo using mouse spinal cord sections.

Read More
Answer

That's great news! Thanks so much for letting us know.

Please feel free to contact me if you need anything further.

Read More

Question

I have used this antibody on astrocytes grown in culture in ICC and mousespinal cord via IHC-Fr. In neither application did I see any signal using 1:200 dilution of the antibody; using another antibody to an astrocyte marker protein, GFAP, is was able to detect astrocytes in both applications using the same samples.

Read More
Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1080427.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Immunohistochemistry (Frozen sections) abreview for Anti-S100 antibody [4C4.9] - Astrocyte Marker

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (skin)
Specification
skin
Fixative
Acetone
Permeabilization
Yes - 0,1% Saponin / PBS
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Feb 27 2012

Question

Thanks for the antibody. It works fine the immunostaining. But I still can not get any signal in western blot, although it was reported that it should work in western blot.



Do you have any other recommendations for western blot?

Read More
Answer

Thank you for your reply. After reviewing the customer feedback for ab4066, it appears that the customers who had obtained good WB results were using BSA as a blocking agent. The feedback we had from a customer using 5% milk was that their WB was unsuccessful. It is possible that this antibody may be sensitive to milk and may produce better results when using BSA.

For both ab4066 andab868, you could also try increasing the primary antibody concentration to the range 1:200-1:500 or loading more protein (30-50ug per lane). I hope this helps, please let me know if you need any further assistance.

Read More

Question

I am trying to do Western Blot and ICC for s100, a marker for astrocytes. The antibody which I got from Abcam is ab4066. I did not get any signal for WB and very weak signal for ICC. So I wonder what is the problem. If I still can not get any signal after trouble shooting, which antibodies do you recommend for WB and ICC, respectively?

Read More
Answer

Thank you for contacting us. I am sorry that ab4066 was not producing satisfactory signal in ICC and WB. Our antibody ab868 is rated highly for use in ICC and WB on human, mouse, rat, cow, and pig samples. If you could please provide some additional details about the protocol and samples you are using, I would be happy to assist you further. If ab4066 is not working as stated on the datasheet and you have purchased it in the past six months, I would be happy to send you ab868 as a free of charge replacement. Could you please provide the following information? 1) What is your original order or PO number? 2) What species and sample type were you testing? How were the cells / tissuefixed or lysed? 3) What permeabilization and/ or blocking agents were used? 4) Could you please tell me the dilution and incubation times for the primary and secondary antibodies? 5) It would be very helpful if you could please provide an image of your WB or ICC.

Read More

1-10 of 21 Abreviews or Q&A

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