Recombinant human GCSF Receptor protein (Fc Chimera) (ab83994)

Description

  • Product name

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

    The ED50 of ab83994 is typically 2-3 ng/ml as measured by its ability to neutralize GCSF mediated proliferation of the murine myeloblastic m-NFS-60 cell line.
  • Purity

    > 95 % SDS-PAGE.

  • Expression system

    HEK 293 cells
  • Accession

  • Protein length

    Protein fragment
  • Animal free

    No
  • Nature

    Recombinant
    • Species

      Human
    • Sequence

      Theoretical Sequence: ECGHISVSAPIVHLGDPITASCIIKQNCSHLDPEPQILWRLGAELQPG GRQQRLSDGTQESIITLPHLNHTQAFLSCCLNWGNSLQILDQVELRAG YPPAIPHNLSCLMNLTTSSLICQWEPGPETHLPTSFTLKSFKSRGNCQ TQGDSILDCVPKDGQSHCCIPRKHLLLYQNMGIWVQAENALGTSMSPQ LCLDPMDVVKLEPPMLRTMDPSPEAAPPQAGCLQLCWEPWQPGLHINQ KCELRHKPQRGEASWALVGPLPLEALQYELCGLLPATAYTLQIRCIRW PLPGHWSDWSPSLELRTTERAPTVRLDTWWRQRQLDPRTVQLFWKPVP LEEDSGRIQGYVVSWRPSGQAGAILPLCNTTELSCTFHLPSEAQEVAL VAYNSAGTSRPTPVVFSESRGPALTRLHAMARDPHSLWVGWEPPNPWP QGYVIEWGLGPPSASNSNKTWRMEQNGRATGFLLKENIRPFQLYEIIV TPLYQDTMGPSQHVYAYSQEMAPSHAPELHLKHIGKTWAQLEWVPEPP ELGKSPLTHYTIFWTNAQNQSFSAILNASSRGFVLHGLEPASLYHIHL MAASQAGATNSTVLTLMTLTPRSSNTKVDKKVEPKSCDKTHTCPPCPA PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA LPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGK
    • Amino acids

      25 to 621
    • Additional sequence information

      Encodes the signal peptide and extracellular domain of human G-CSF R (aa 1-621) was fused to the Fc region of human IgG1 (aa 90-330). The chimeric protein was expressed in modified human 293 cells.

Specifications

Our Abpromise guarantee covers the use of ab83994 in the following tested applications.

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

  • Applications

    SDS-PAGE

    Functional Studies

  • Form

    Lyophilised
  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. After reconstitution store at -20ºC. Avoid freeze / thaw cycles.

    Constituents: 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

    • CD 114
    • CD114
    • CD114 antigen
    • Colony stimulating factor 3 receptor
    • Colony stimulating factor 3 receptor (granulocyte)
    • CSF 3R
    • CSF3R
    • CSF3R_HUMAN
    • Csfgr
    • G CSF R
    • G-CSF receptor
    • G-CSF-R
    • GCSFR
    • Granulocyte colony stimulating factor receptor
    • Granulocyte colony-stimulating factor receptor
    • OTTHUMP00000009703
    • OTTHUMP00000009704
    • OTTHUMP00000009705
    see all
  • Function

    Receptor for granulocyte colony-stimulating factor (CSF3), essential for granulocytic maturation. Plays a crucial role in the proliferation, differientation and survival of cells along the neutrophilic lineage. In addition it may function in some adhesion or recognition events at the cell surface.
  • Tissue specificity

    One or several isoforms have been found in myelogenous leukemia cell line KG-1, leukemia U-937 cell line, in bone marrow cells, placenta, and peripheral blood granulocytes. Isoform GCSFR-2 is found only in leukemia U-937 cells. Isoform GCSFR-3 is highly expressed in placenta.
  • Involvement in disease

    Hereditary neutrophilia
  • Sequence similarities

    Belongs to the type I cytokine receptor family. Type 2 subfamily.
    Contains 5 fibronectin type-III domains.
    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
  • Domain

    The WSXWS motif appears to be necessary for proper protein folding and thereby efficient intracellular transport and cell-surface receptor binding.
    The box 1 motif is required for JAK interaction and/or activation.
  • Cellular localization

    Secreted and Cell membrane.
  • Information by UniProt

Images

  • 1D SDS-PAGE of ab83994 before and after treatment with glycosidases to remove oligosaccharides.
    Lane 1: ab83994
    Lane 2: ab83994 treated with PNGase F to remove potential N-linked glycans
    Lane 3: ab83994 treated with a glycosidase cocktail to remove potential N- and O-linked glycans.
    Approximately 5 μg of protein was loaded per lane; Gel was stained using Coomassie.

    Drop in MW after treatment with PNGase F indicates presence of N-linked glycans. A tightening of the band after treatment with the glycosidase cocktail indicates that O-linked glycans may be present. Additional bands in lane 2 and lane 3 are glycosidase enzymes.

References

ab83994 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer


Each separate size is filled separately so that the target amount of protein of the primary product is maintained.
When each is brought to 0.5ml, each will have only 1% HSA and 10% sugar as a stabilizer. The 25ug size will be 25ug/500ul and the 50ug will be 50ug/500ul, the amount of sugar and HSA is constant in both but the target protein is less in the 25ug size obviously, since the "mass" of the target is matched to the size run.
You can combine both formats into a single stock and you would have approximately 75ug/1mL of target protein that way, probably much easier to figure and work with than changing between the two sizes.

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Answer


"We have our specification that was used to pass the current batch of material. The material would not have been released if it had not worked at the time of release and I've never had any complaints or reason to suspect otherwise.
Our specification is:
The ED50 of G-CSF R – Fc Chimera is typically 2-3 ng/ml as measured by its ability to neutralize G-CSF mediated proliferation of the murine myeloblastic m-NFS-60 cell line.
The chimera's are tested only by their ability to block G-CSF stimulation --note we are using our own G-CSF in the assay, other manufacture's materials may require different amounts of the chimera."

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Answer


The source of the GCSF Receptor protein ab83994 informed me that :
With regards to any of our proteins actually binding HSA, no we do not have any data, and I suppose one could get some minor binding just due to protein-protein interactions since the amount of HSA is rather high and it is there to stabilize the protein but I do not put that sort of interaction in the same class as say a true "binding" where there is a specific avidity for HSA. But we've never examined it and so far it certainly hasn't been a huge factor in the activity assays for any of the proteins we offer.

Now as for dimerization, all we know is that our Fc fusions do what they are supposed to do.
G-CSF R is produced as an ECD-Fc fusion protein with the aim of enhancing its ativity. ECD-Fc fusion proteins have an advantage over soluble receptors because many receptors are only functional in dimeric form. Fusion to the Fc domain of IgG1 induces dimerization due to the ability of the Fc domain to form disulfide bonds. The resulting dimeric receptor ECD-Fc mimics the activated form of the receptor and possess enhanced affinity for its cognate ligand relative to its monomeric form. For a comprehensive review please see Avalos BR (1996) Blood 88(3): 761-777.

With this particular product we test only the the ability of the fusion protein to bind the target and inhibit proliferation, since it does that we are confident that the ECD domain is intact; we have not compared that activity to non-Fc fusion forms so we cannot truly say if the Fc is providing additional efficacy but studying the literature our IgG domain should function as expected so we would expect so. Unfortunately we do not have any data or graphs available.

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Answer

Thank you for your inquiry.

I can confirm that 10% Trehalose and 1% Human serum albumin are contained in the lyophilized product.

Please follow the instructions on the datasheet for reconstitution.

I hope this information is helpful and wish you good luck with your research.

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