Overview

  • Product name
    Anti-S100 antibody [4C4.9]
    See all S100 primary antibodies
  • Description
    Mouse monoclonal [4C4.9] to S100
  • Tested applications
    Suitable for: ICC/IF, IHC-P, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Tissue, cells or virus. 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
  • 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

Anti-S100 antibody [4C4.9] images

  • ab4066 at a dilution of 1/200, staining S100 in astrocytes (Alexa 488 secondary at 1/2000) on 30µm coronal rat brain tissue sections in free floating IHC (see protocol link for detailed description). Image shows astrocytes observed with 20x objective. Image coloured in Photoshop.

    NB: No labeling observed following omission of primary antibody.

    Sections were viewed using an Axioplan 2 Imaging microscope (Imaging Associates) fitted with 10x, 20x and 40x Plan-Neofluorobjectives (Zeiss, Germany) and images were taken using a AxioCam Hrm digital camera (Zeiss, Germany) and AxioVision software (Imaging Associates).

  • 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

  • All lanes : Anti-S100 antibody [4C4.9] (ab4066) at 1/200 dilution

    Lane 1 : 40ug Rat spinal cord homogenate
    Lane 2 : 40ug Rat spinal cord homogenate
    Lane 3 : 40ug Rat spinal cord homogenate
    Lane 4 : 40 ug Mouse spinal cord homogenate
    Lane 5 : 40 ug Mouse spinal cord homogenate
    Lane 6 : 40 ug Mouse spinal cord homogenate
    Lane 7 : 40 ug Rat brain homogenate
    Lane 8 : 40 ug Rat brain homogenate

    Secondary
    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

    This image is courtesy of an Abreview submitted by Ms Nancy Nutile-McMenemy

    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

  • ab4066 at a 1/100 dilution staining rat cortical preps grown 14 days in culture, plated onto coverslips. The preps were acid/alcohol fixed for 15 minutes, blocked and then incubated with the antibody for 1 hour. Bound antibody was detected using a Alexa fluor ® 555 conjugated goat polyclonal antibody. Nuclei were visualised using DAPI (blue). Note: s100 is colocalizing around the nucleus.

    See Abreview

  • ab4066 staining human melanoma by IHC-P.

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

This product has been referenced in:
  • Platt RJ  et al. Chd8 Mutation Leads to Autistic-like Behaviors and Impaired Striatal Circuits. Cell Rep 19:335-350 (2017). Read more (PubMed: 28402856) »
  • Raha-Chowdhury R  et al. Expression and cellular localization of hepcidin mRNA and protein in normal rat brain. BMC Neurosci 16:24 (2015). IHC . Read more (PubMed: 25896789) »

See all 23 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

Submitted Aug 19 2016

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
Username

Abcam user community

Verified customer

Submitted Dec 14 2015

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
Username

Abcam user community

Verified customer

Submitted May 14 2015


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 mu...

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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 n...

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

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

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 con...

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Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Feb 27 2012

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 unsuc...

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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 de...

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1-10 of 21 Abreviews or Q&A

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