Monoclonal mouse anti human G-CSF

///Monoclonal mouse anti human G-CSF

Monoclonal mouse anti human G-CSF

$99.00$149.00


accession P09919


Applications

ELISA: This antibody can be used as capture antibody at 2 μg/mL jointly with biotinylated Anti GCSF polyclonal detection antibody to detect Human GCSF. The detection limit for recombinant Human G-CSF is approximately 0.1 ng/well.

Western blot:This monoclonal antibody has been tested in WB analysis of human G-CSF . A suitable range of concentrations of this antibody for WB detection is 1-2 µg/ml.
The detection limit for Recombinant Human G-CSF is 1.0-2.0 ng/lane, under reducing or non-reducing conditions.

Source This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with recombinant E. coli-derived Human GCSF.
Species reactivity Human G-CSF
Purification The IgG fraction of the tissue culture supernatant was purified by Protein A affinity chromatography.
Presentation Prepared from sterile filtered solution at a concentration of 1mg/ml in PBS with 5% trehalose.
Reconstitution A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers
Storage This antibody can be stored in working aliquots at 2° - 8° C for one month, or at -20° C for six months without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage This antibody product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.


Interactor P40223
Molecular function Cytokine
Molecular function Growth-factor

Methods

In vivo tumor growth and G-CSF studies

  • Female BALB/c mice were injected orthotopically in an abdominal mammary gland with 5×104 4T1 cells.
  • Female B6 mice were injected similarly with 5×105 AT-3 cells, including those cell lines used in the knockdown studies.
  • For the G-CSF over-expression tumor studies, syngeneic BALB/c mice were injected subcutaneously into the flank with 4×105 CMS4 cells.
  • In the admix experiments, 5×104 4T1 tumor cells were mixed with MACS-purified, MHC-matched CD11b+Gr-1+ cells at a 2∶1 ratio, respectively, prior to orthotopic implantation.
  • Spleens, serum and tumor tissue were recovered, as appropriate.
  • For the G-CSF blockade studies, B6 mice were injected orthotopically, and on day 7 when tumors became palpable, mice were injected intraperitoneally with 10 µg of either neutralizing anti-G-CSF mAb or rat IgG isotype for 8-consecutive days, as described in vivo depletion experiments, BALB/c mice were injected ip with anti-Gr-1 Ab (cloneB6-8C5, , ) or control Ab (at IgG2b clone LTF-2,…
  • Female BALB/c mice were injected orthotopically in an abdominal mammary gland with 5×104 4T1 cells.
  • Female B6 mice were injected similarly with 5×105 AT-3 cells, including those cell lines used in the knockdown studies.
  • For the G-CSF over-expression tumor studies, syngeneic BALB/c mice were injected subcutaneously into the flank with 4×105 CMS4 cells.
  • In the admix experiments, 5×104 4T1 tumor cells were mixed with MACS-purified, MHC-matched CD11b+Gr-1+ cells at a 2∶1 ratio, respectively, prior to orthotopic implantation.
  • Spleens, serum and tumor tissue were recovered, as appropriate.
  • For the G-CSF blockade studies, B6 mice were injected orthotopically, and on day 7 when tumors became palpable, mice were injected intraperitoneally with 10 µg of either neutralizing anti-G-CSF mAb or rat IgG isotype for 8-consecutive days, as described in vivo depletion experiments, BALB/c mice were injected ip with anti-Gr-1 Ab (cloneB6-8C5, , ) or control Ab (at IgG2b clone LTF-2, , ).
  • Antibodies were administered at 200 µg/mouse once tumors became palpable, and continued twice weekly for the duration of the experiment, similarly as described 3 or for other health considerations.
  • Tumor growth was measured 2 – 3 times/week in two dimensions and tumor volume was calculated using the formula (width2×length)/2.
  • For recombinant G-CSF in vivo studies, mice were injected subcutaneously with recombinant G-CSF (10 µg/day, , ) for 5 consecutive days and serum/tissues collected, similarly as described

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