Human Anti-MET Recombinant Antibody (clone 5D5) (CAT#: PABL-271)

Recombinant Human Antibody (5D5) is capable of binding to MET, expressed in HEK 293 cells. Expressed as the combination of a heavy chain (HC) containing VH from anti-MET mAb and CH1-3 region of human IgG1 and a light chain (LC) encoding VL from anti-MET mAb and CL of human light chain. Exists as a disulfide linked dimer of the HC and LC hetero-dimer under non-reducing condition. This antibody potently inhibits HGF binding and receptor phosphorylation and signaling and has antibody-like pharmacokinetics and antitumor activity.

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  • Published Data
  • Tested Data
  • Gene Expression
  • Datasheet
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  • COA

Figure 1 MKN45 cells treated with 5 μg/ml of F46 or 5D5 were incubated for 24 h. c-Met degradation was measured by Western blot (A) and ELISA (B). Phosphorylation of c-Met and Akt in Caki-1 cells was measured by Western blot (C, E) and ELISA (D, F). Caki-1 cells were incubated for 30 min with 5 μg/ml of F46 or 5D5 or 100 ng/ml of HGF. The error bars indicate the mean ± SD of three independent experiments. Phosphorylation of c-Met downstream kinases in Caki-1 (G) and NCI-H441 (I) cells were measured by Western blot. Co-immunoprecipitation experiment showed induced binding between Gab1 and c-Met by 5D5, but not by F46 (H).

Figure 1 MKN45 cells treated with 5 μg/ml of F46 or 5D5 were incubated for 24 h. c-Met degradation was measured by Western blot (A) and ELISA (B). Phosphorylation of c-Met and Akt in Caki-1 cells was measured by Western blot (C, E) and ELISA (D, F). Caki-1 cells were incubated for 30 min with 5 μg/ml of F46 or 5D5 or 100 ng/ml of HGF. The error bars indicate the mean ± SD of three independent experiments. Phosphorylation of c-Met downstream kinases in Caki-1 (G) and NCI-H441 (I) cells were measured by Western blot. Co-immunoprecipitation experiment showed induced binding between Gab1 and c-Met by 5D5, but not by F46 (H).

Oh, Y. M., Song, Y. J., Lee, S. B., Jeong, Y., Kim, B., Kim, G. W.,... & Nam, D. H. (2012). A new anti-c-Met antibody selected by a mechanism-based dual-screening method: therapeutic potential in cancer. Molecules and cells, 34(6), 523-529.

Figure 2 MKN45 cells were incubated with increasing amounts of recombinant human HGF. They were added with 0.5 μg of 5D5 or F46 followed by flow cytometry. ΔMFI (y-axis) indicates the decrease in percentage of mean fluorescence intensity (MFI) upon treatment with F46 or 5D5 (top). For the competition ELISA, biotinylated HGF and differing concentrations of IgG, F46, or free HGF were treated on c-Met coated plate (bottom).

Figure 2 MKN45 cells were incubated with increasing amounts of recombinant human HGF. They were added with 0.5 μg of 5D5 or F46 followed by flow cytometry. ΔMFI (y-axis) indicates the decrease in percentage of mean fluorescence intensity (MFI) upon treatment with F46 or 5D5 (top). For the competition ELISA, biotinylated HGF and differing concentrations of IgG, F46, or free HGF were treated on c-Met coated plate (bottom).

Oh, Y. M., Song, Y. J., Lee, S. B., Jeong, Y., Kim, B., Kim, G. W.,... & Nam, D. H. (2012). A new anti-c-Met antibody selected by a mechanism-based dual-screening method: therapeutic potential in cancer. Molecules and cells, 34(6), 523-529.

Figure 3 A549, H441, and MDA-MB-435 cells were kept serum-free and treated with 10 μg/ml rSema or anti-Met 5D5-Fab.

Figure 3 A549, H441, and MDA-MB-435 cells were kept serum-free and treated with 10 μg/ml rSema or anti-Met 5D5-Fab.

The samples were immunoblotted with phospho-tyrosine, Met, phospho-MAPK, or MAPK antibodies.

Kong-Beltran, M., Stamos, J., & Wickramasinghe, D. (2004). The Sema domain of Met is necessary for receptor dimerization and activation. Cancer cell, 6(1), 75-84.

Figure 4 293 cells were transfected with the indicated constructs and analyzed by 4%–12% SDS-PAGE. Lysates were immunoprecipitated with anti-Met 5D5 antibody and immunoblotted with His antibody (top) and reprobed with Met C-12 antibody (bottom).

Figure 4 293 cells were transfected with the indicated constructs and analyzed by 4%–12% SDS-PAGE. Lysates were immunoprecipitated with anti-Met 5D5 antibody and immunoblotted with His antibody (top) and reprobed with Met C-12 antibody (bottom).

Kong-Beltran, M., Stamos, J., & Wickramasinghe, D. (2004). The Sema domain of Met is necessary for receptor dimerization and activation. Cancer cell, 6(1), 75-84.

Figure 5 rSema and anti-Met 5D5-Fab inhibit cell motility.

Figure 5 rSema and anti-Met 5D5-Fab inhibit cell motility.

A: H441 cells were scraped on Day 0 and treated with increasing concentrations of rSema plus HGF in serum-free media. The scrapes were photographed on Day 0 and Day 1. A representative data set of three independent experiments is shown. B: H441 cells were scraped and treated with mock, rSema, or anti-Met 5D5-Fab in serum-free media over the course of 2 days. Photographs were taken on Day 0 and Day 2.

Kong-Beltran, M., Stamos, J., & Wickramasinghe, D. (2004). The Sema domain of Met is necessary for receptor dimerization and activation. Cancer cell, 6(1), 75-84.

Figure 6 Cell migration is inhibited by rSema and anti-Met 5D5-Fab.

Figure 6 Cell migration is inhibited by rSema and anti-Met 5D5-Fab.

Kong-Beltran, M., Stamos, J., & Wickramasinghe, D. (2004). The Sema domain of Met is necessary for receptor dimerization and activation. Cancer cell, 6(1), 75-84.


Specifications

  • Immunogen
  • The details of the immunogen for this antibody are not available.
  • Host Species
  • Human
  • Derivation
  • Human
  • Type
  • Human IgG
  • Specificity
  • Human MET
  • Species Reactivity
  • Human
  • Clone
  • 5D5
  • Applications
  • WB, Block, FuncS

Product Property

  • Purity
  • >95% as determined by SDS-PAGE and HPLC analysis
  • Concentration
  • Please refer to the vial label for the specific concentration.
  • Buffer
  • PBS
  • Preservative
  • No preservatives
  • Storage
  • Centrifuge briefly prior to opening vial. Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.

Target

  • Alternative Names
  • MET; MET proto-oncogene; receptor tyrosine kinase; HGFR; AUTS9; RCCP2; c-Met; hepatocyte growth factor receptor; SF receptor; HGF receptor; HGF/SF receptor; proto-oncogene c-Met; scatter factor receptor; tyrosine-protein kinase Met; met proto-oncogene tyrosine kinase

Related Resources

  • Related Diseases

Product Notes

This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:

• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production

See more details about Hi-Affi™ recombinant antibody benefits.

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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.

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