Anti-Human CXCR4 Therapeutic Antibody (Ulocuplumab) (CAT#: TAB-H72)

Recombinant human antibody to Human CXCR4

  • Published Data
  • Datasheet
  • MSDS
  • COA

Figure 1 SIA induction by Ulocuplumab (BMS-936564) is CXCR4 specific as observed in Ramos and primary CLL cells but not in K562.

Ramos and K562 cell lines were treated with different concentrations of Ulocuplumab (BMS-936564) and AMD3100 for 48 hrs followed by running the samples on flow cytometer to study the % SIA. Treatment with AMD3100 in Ramos/ K562/CLL did not induce significant % SIA as compared to Ulocuplumab (BMS-936564). In contrast, Ulocuplumab (BMS-936564) induced significant % SIA in Ramos and CLL cells (CXCR4+), but not in K562 (CXCR4-) as evaluated by flow cytometry).

Kashyap, M. K. , Kumar, D. , Jones, H. , Amayachanaga, C. I. , Choi, M. Y. , & Melocardenas, J. , et al. (2016). Ulocuplumab (bms-936564 / mdx1338): a fully human anti-cxcr4 antibody induces cell death in chronic lymphocytic leukemia mediated through a reactive oxygen species-dependent pathway. Oncotarget, 7(3), 2809-2822.

Figure 2 SIA induction by Ulocuplumab (BMS-936564) is CXCR4 specific as observed in Ramos and primary CLL cells but not in K562.

The CLL cells cultured either alone or with stromal cell support were treated with Ulocuplumab (BMS-936564) or AMD3100 for 48 hrs at 37 o C followed by flow cytometer for measurement of % SIA. AMD3100 does not induce significant % SIA in CLL as compared to Ulocuplumab (BMS-936564). Ulocuplumab (BMS-936564) induced significant amount of % SIA in CLL cells alone or CLL co-cultured with stromal cell support.

Kashyap, M. K. , Kumar, D. , Jones, H. , Amayachanaga, C. I. , Choi, M. Y. , & Melocardenas, J. , et al. (2016). Ulocuplumab (bms-936564 / mdx1338): a fully human anti-cxcr4 antibody induces cell death in chronic lymphocytic leukemia mediated through a reactive oxygen species-dependent pathway. Oncotarget, 7(3), 2809-2822.

Figure 3 Ulocuplumab Inhibits the EMTPhenotype Acquisition in MM Cells BothIn Vitro and In Vivo.

MM.1S cells were cultured in presenceor absence of ulocuplumab (0.025–1 mM; 6 hr).MM.1S cells were then harvested, and cell lysateswere subjected to western blot using anti-p-b-catenin,-b-catenin, -p-GSK3-b, -tubulin, -Snail,-Twist, -Slug, and -E-cadherin antibodies. Adjustedband intensity for each protein normalized to therelative loading control is provided within FiguresS5A and S5B and expressed as fold of control(control corresponds to untreated cells).

Roccaro, A. , Mishima, Y. , Sacco, A. , Moschetta, M. , Tai, Y. T. , & Shi, J. , et al. (2015). Cxcr4 regulates extra-medullary myeloma through epithelial-mesenchymal-transition-like transcriptional activation. Cell Reports, 12(4), 622-635.

Figure 4 Ulocuplumab Inhibits the EMTPhenotype Acquisition in MM Cells BothIn Vitro and In Vivo.

Bone chips from donor mice wereloaded with CXCR4+ MM.1S and implanted sub-cutaneously into SCID/Bg-recipient mice. Micewere treated with either control Ab or ulocuplumab(n = 5/group; 10 mg/kg; i.p.; 43to 53/week).Ulocuplumab led to inhibited MM cell growthwithin the implanted bone (implanted bone isshown in C), together with inhibited ability of MMcells to metastatize from bone to bone (host boneis shown in D).

Roccaro, A. , Mishima, Y. , Sacco, A. , Moschetta, M. , Tai, Y. T. , & Shi, J. , et al. (2015). Cxcr4 regulates extra-medullary myeloma through epithelial-mesenchymal-transition-like transcriptional activation. Cell Reports, 12(4), 622-635.

Figure 5 Ulocuplumab Inhibits the EMTPhenotype Acquisition in MM Cells BothIn Vitro and In Vivo.

The ability of ulocuplumab to modulate EMTwas demonstrated ex vivo: decreased mRNAlevels of Twist, Snail, and Slug, together withincreased mRNA levels of E-cadherin, wereobserved in BM cells obtained from the host fe-murs of ulocuplumab-treated mice. No MM cell-injected mice were used as control. p indicates pvalue. Average of experiments performed in tripli-cate is shown. Error bars indicate SD.

Roccaro, A. , Mishima, Y. , Sacco, A. , Moschetta, M. , Tai, Y. T. , & Shi, J. , et al. (2015). Cxcr4 regulates extra-medullary myeloma through epithelial-mesenchymal-transition-like transcriptional activation. Cell Reports, 12(4), 622-635.

Figure 6 Ulocuplumab Exerts Anti-MMActivity, Either as Single Agent or in Combi-natory Regimens In Vivo, and InducesToxicity on Primary MM Cells.

SCID/Bg mice were injected with 5 3106MM.1S-GFP+/Luc+cells i.v. and treated with ulo-cuplumab (10 mg/kg; 33to 43/week; i.p.) or iso-type control antibody (30 mg/kg; 33to 43/week;i.p.). Ulocuplumab led to inhibited MM cell homingto the BM, as shown by using intravital confocalmicroscopy at the 3rdweek. (GFP+MM cells, greencolor; Evans-Blue-positive blood vessels, redcolor). High-resolution images with cellular detailwere obtained through the intact mouse skull atdepths of up to 250 mm from the surface of the skullusing a 1030.45NA Plan-Apo objective (CarlZeiss) and assembled together to generate a finalimage that depicts the whole bone marrow.

Roccaro, A. , Mishima, Y. , Sacco, A. , Moschetta, M. , Tai, Y. T. , & Shi, J. , et al. (2015). Cxcr4 regulates extra-medullary myeloma through epithelial-mesenchymal-transition-like transcriptional activation. Cell Reports, 12(4), 622-635.

Figure 7 Ulocuplumab Exerts Anti-MMActivity, Either as Single Agent or in Combi-natory Regimens In Vivo, and InducesToxicity on Primary MM Cells.

BM-derived primary CD138+ cells werecultured in presence or absence of ulocuplumab(50–400 nM; 48 hr). Cell toxicity was performed byusing MTT. Ulocuplumab exerted anti-MM activityagainst primary MM cells. Error bars indicate SD.

Roccaro, A. , Mishima, Y. , Sacco, A. , Moschetta, M. , Tai, Y. T. , & Shi, J. , et al. (2015). Cxcr4 regulates extra-medullary myeloma through epithelial-mesenchymal-transition-like transcriptional activation. Cell Reports, 12(4), 622-635.


Specifications

  • Host Species
  • human
  • Derivation
  • human
  • Type
  • IgG4 - kappa
  • Specificity
  • CXCR4(chemokine (C-X-C motif) receptor 4, fusin, stromal cell-derived factor 1 receptor, SDF-1 receptor, CXCL12 receptor, CD184) [Homo sapiens]
  • Species Reactivity
  • Human
  • Applications
  • IP, IF, FuncS, FC, Neut, ELISA, ICC
  • CAS
  • 1375830-34-4
  • Generic Name
  • ulocuplumab
  • MW
  • 146.2 kDa
  • Related Disease
  • Acute myeloid leukemia (AML)

Applications

  • Application Notes
  • The CXCR4 antibody has been reported in applications of FC, Cyt, WB, IHC, RT-PCR, IF, MTT.
    WB: Cells were harvested and lysed using lysis buffer supplemented with 5 mM NaF, 2 mM Na3VO4, 1 mM PMSF (polymethylsulfonylfluoride), 5 mg/ml leupeptin, and 5 mg/ml aprotinin.
    IHC: Autopsy specimens of synchronous secondary extramedullary MM sites wereanalyzed for the expression of CD138 and CXCR4. Murine BM tissues (femurs)were analyzed for the expression of human CD138 and stained with hema-toxylin-eosin.

Target

  • Alternative Names
  • ulocuplumab;1375830-34-4;MDX-1338;BMS-936564;CXCR4;chemokine (C-X-C motif) receptor 4;chemokine (C X C motif), receptor 4 (fusin);C-X-C chemokine receptor type 4;CD184;D2S201E;fusin;HM89;HSY3RR;LESTR;NPY3R;NPYR;NPYY3R;CXC-R4;CXCR-4;CD184 antigen;SDF-1 rec

For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

Related Resources

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* Abbreviations
3D IHC3D Immunohistochemistry
ActivActivation
AgonistAgonist
ApopApoptosis
BABioassay
BIBioimaging
BlockBlocking
Cell ScreeningCell Screening
SeparationCell Separation
ChIPChromatin Immunoprecipitation
CMCDComplement Mediated Cell Depletion
CostimCostimulation
CytCytotoxicity
DepletionDepletion
DBDot Blot
EMElectron Microscopy
ELISAEnzyme-linked Immunosorbent Assay
ELISPOTEnzyme-linked Immunosorbent Spot
FCFlow Cytometry
FuncSFunctional Assay
GSGel Super Shift Assay
HAHemagglutination
IAImmunoassay
IBImmunoblotting
ICCImmunocytochemistry
IDImmunodiffusion
IFImmunofluorescence
IHCImmunohistochemistry
IHC-FrImmunohistochemistry-Frozen
IHC-PImmunohistochemistry-Paraffin
REImmunohistology - Resin Sections
IPImmunoprecipitation
IRMAImmunoradiometric Assay
SHIn situ hybridization
InhibInhibition
ICFCIntracellular Staining for Flow Cytometry
KO/KD-WBKnockout/Knockdown target confirmation by Western Blot
Live cell imagingLive cell imaging
CyTOF®Mass Cytometry
MeDIPMethylated DNA Immunoprecipitation
MultiplexMultiplex bead-based assay
NeutNeutralization
PPProtein Purification
PGProteogenomics
RIRadial Immunodiffusion
RIARadioimmunoassay
StimStimulation
SPRSurface Plasmon Resonance
TCTissue Culture
TBTurbidimetry
WBWestern Blot

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