Recombinant human MCSF Receptor protein (Fc Chimera) (ab83682)

Description

  • Product name

    Recombinant human MCSF Receptor protein (Fc Chimera)
    See all MCSF Receptor proteins and peptides
  • Biological activity

    Activity: The ED50 of ab83682 is typically 0.2 - 0.3 ug/ml as measured by its ability to neutralize MCSF mediated proliferation of mNFS-60 cells.
  • Purity

    > 95 % SDS-PAGE.

  • Expression system

    HEK 293 cells
  • Accession

  • Protein length

    Protein fragment
  • Animal free

    No
  • Nature

    Recombinant
    • Species

      Human
    • Sequence

      Theoretical Sequence: IPVIEPSVPELVVKPGATVTLRCVGNGSVEWDGPPSPHWTLYSDGSSSIL STNNATFQNTGTYRCTEPGDPLGGSAAIHLYVKDPARPWNVLAQEVVVFE DQDALLPCLLTDPVLEAGVSLVRVRGRPLMRHTNYSFSPWHGFTIHRAKF IQSQDYQCSALMGGRKVMSISIRLKVQKVIPGPPALTLVPAELVRIRGEA AQIVCSASSVDVNFDVFLQHNNTKLAIPQQSDFHNNRYQKVLTLNLDQVD FQHAGNYSCVASNVQGKHSTSMFFRVVESAYLNLSSEQNLIQEVTVGEGL NLKVMVEAYPGLQGFNWTYLGPFSDHQPEPKLANATTKDTYRHTFTLSLP RLKPSEAGRYSFLARNPGGWRALTFELTLRYPPEVSVIWTFINGSGTLLC AASGYPQPNVTWLQCSGHTDRCDEAQVLQVWDDPYPEVLSQEPFHKVTVQ SLLTVETLEHNQTYECRAHNSVGSGSWAFIPISAGAHTHPPDEGSSNTKV DKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
    • Amino acids

      1 to 512
    • Additional sequence information

      Fused with the Fc region of Human IgG1 at the C-terminus.

Specifications

Our Abpromise guarantee covers the use of ab83682 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • Applications

    Functional Studies

    SDS-PAGE

  • Form

    Lyophilised
  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Store at +4°C.

    Constituents: PBS, 1% Human serum albumin, 10% Trehalose

    This product is an active protein and may elicit a biological response in vivo, handle with caution.

  • Reconstitution
    It is recommended that 0.5 ml of sterile phosphate-buffered saline be added to the vial. Following reconstitution short-term storage at 4°C is recommended and longer-term storage of aliquots at -18 to -20°C. Repeated freeze thawing is not recommended.

General Info

  • Alternative names

    • C FMS
    • CD 115
    • CD115
    • CD115 antigen
    • CFMS
    • Colony stimulating factor 1 receptor
    • Colony stimulating factor I receptor
    • CSF 1 R
    • CSF 1R
    • CSF-1 receptor
    • CSF-1-R
    • CSF1 R
    • CSF1R
    • CSF1R_HUMAN
    • CSFR
    • EC 2.7.10.1
    • FIM 2
    • FIM2
    • FMS
    • FMS proto oncogene
    • FMS protooncogene
    • HDLS
    • M-CSF Receptor
    • M-CSF-R
    • Macrophage colony stimulating factor 1 receptor
    • Macrophage colony stimulating factor I receptor
    • Macrophage colony-stimulating factor 1 receptor
    • McDonough feline sarcoma viral (v fms) oncogene homolog
    • MCSFR
    • Oncogen FMS
    • Proto-oncogene c-Fms
    • V-FMS McDonough feline sarcoma viral oncogen homolog, formerly
    see all
  • Function

    Protein tyrosine-kinase transmembrane receptor for CSF1 and IL34.
  • Tissue specificity

    Expressed in bone marrow and in differentiated blood mononuclear cells.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily.
    Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 1 protein kinase domain.
  • Cellular localization

    Membrane.
  • Information by UniProt

Images

  • Densitometry of protein isoforms visualised by 2-DE. The triangle indicates the theoretical MW and pI of the protein.
  • 1D SDS-PAGE of ab83682 before and after treatment with glycosidases to remove oligosaccharides.
    Lane 1 MW markers; Lane 2 ab83682; Lane 3 ab83682 treated with PNGase F to remove potential N-linked glycans; Lane 4 ab83682 treated with a glycosidase cocktail to remove potential N- and O-linked glycans. Approximately 5 μg of protein was loaded per lane.

    Drop in MW after treatment with PNGase F indicates presence of N-linked glycans. A further drop in MW after treatment with the glycosidase cocktail indicates the presence of O-linked glycans. Additional bands in lane 3 and lane 4 are glycosidase enzymes.

  • A sample of ab83682 without carrier protein was reduced and alkylated and focused on a 3-10 IPG strip then run on a 4-20% Tris-HCl 2D gel. Approximately 40 μg of protein was load; Gel was stained using Deep Purple™.

References

ab83682 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for contacting me.

I received an answer from the lab. The lab confirmed, that for the ab83682 a NuPage 4-20% Tris-Glycine SDS gel was used together with a standard buffer system.

The lab also confirmed, that it is not recommended to use the protein in a native PAGE. They would expect different bands in a native Page compared to the SDS-PAGE.

I hope this information might be helpful for you. Do not hesitate to contact me again if you have further questions.

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Answer

Thank you very much for providing your WB result.

1. As far as I can see the band detected in samples 5 to 7 are at the correct size between 100-140 kDa. I think the band that shows up at about 70 kda might still be the BSA. Because of the high amount of BSA protein the antibody might get caught and for that reason a band will appear on the blot.

To demonstrate the specifics of the antibody for the MCSF protein and that this is the band detected between 100-140 kDa I can recommend to have a look at the following protocol:

https://www.abcam.com/index.html?pageconfig=resource&rid=11378

Could you confirm how the expected size for the E. coli derived and the HEK derived protein was validated? Did you contact the supplier to make sure they should be between 100 - 140 kDa as well?

I hope this information will help you and I wish you good luck with the experiment. If you have any further questions do not hesitate to ask again.

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Answer

Thank you for this new image.

It would be beneficial to have a new western blot result to assure which bands are the MCSF protein. Like mentioned before, I suspect the strong band at about 77 kDa might be BSA, which is a content of the storage buffer.

The third band which is detected under denatured conditions might be the protein without modification?

I look forward to hear from you with the new WB result.

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Answer

Thank you for your email.

Thank you for giving me this further information. I am pleased that you will try the given suggestions for the experiment. I hope this might be helpful for you.

I can confirm that I think there are bands at the right size. The smaller bands might be the protein without modification but they also run at the size of the mentioned BSA, which is a component of the storage buffer for the protein.

I look forward to receive your WB results for more confirmation in this case.

I like to suggest to contact the company where you got the enzyme from. If you expect different sizes after digestion with the ChABC they might be able to help you to optimise this part of the protocol.

I look forward hearing from you again and wish you good luck with the experiment.

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Answer

I apologize for making you wait this long. Regrettably I did not receive a confirmation about the used gel from the lab. I wrote them a reminder email as you did with me. Thank you very much for this.

Meanwhile I was looking through the data you kindly provided.

Your protocols are looking very nice and I am sure everything was done thoroughly.

There are a few thinks I would like to ask and to suggest while waiting for an answer of the lab.

1. As far as I can see the gel shows bands between 100 kDa and 140 kDa as well as the observed band at about 70 kDa. I would suggest the dominant band at 70 kDa might be serum albumin which is a component of the buffer for the MCSF protein (Fc Chimera Active) (ab83682) and possibly also for the other used proteins. The additional bands between 100 and 140 kDa seem to be the specific protein.

2. Could you confirm that the samples in lane 2-4 are the MCSF protein (Fc Chimera Active) (ab83682)?

3. In lane 5-7 we would suggest that the loaded amount of protein seems very high and for that reason the observed bands are not very clear. We would suggest reducing the amount of loaded protein in this case.

4. Could you confirm what markers were used for the ponceau staining and the WB? I have never seen a marker before with such a high molecular weight. Furthermore the size seems to be very different to the used marker for the SDS-PAGE.

5. Reviewing the WB result it seems, that the detection of protein is specific. In lane 1-4 are two bands detected. The lower band might be the protein without glycolisation, the higher band the protein with glycolisation. The detected bands in lane 5-7 might be of higher molecular weight because the Fc-fusion tag which would change the molecular weight.

I would like to hear your opinion to these suggestions and hope they might be helpful to you. As soon as I hear back from the lab I will forward the information that I get for you. Please do not hesitate to contact me again with any further questions.

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Question
Answer

I like to reassure you that we have not forgotten about your inquiry. Anja is currently out of office. However, I know about your inquiry and I will get back to you as soon as I receive an answer from the lab. Regrettably I have not received the needed information to help you yet.

Thank you very much for your patience. I will write you again as soon as I get the answer for your question from the lab.

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