Mouse Anti-αV Integrins Recombinant Antibody (clone 17E6) (CAT#: PABX-191)

This product is a recombinant mouse antibody that recognizes human αV Integrins. This antibody is a primate-specific mouse mAb that inhibits V integrins, is in phase II trials for treating cancer.


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Figure 1 The alpha-v group antibodies immunoprecipitate similar proteins.

Figure 1 The alpha-v group antibodies immunoprecipitate similar proteins.

Viable M21 melanoma (A) were surface labelled, extracted with detergent, and immunoprecipitated and resolved on a non-reducing 7.5% SDSPAGE gel. Migration of standards (weights in kDa) are indicated to the left. Precipitation was with the control antibodies LM142 (anti-αv: lane a) and LM609 (anti-αvβ3: lane b), and 21H6 (lane c), 10G2 (lane d), 20A9 (lane e), 23G5 (lane f), 17E6 (lane g), 14D9 (lane h), and 14E2 (lane i). LM142 and LM609 precipitate a similar pattern of proteins to 10G2, 20A9, 23G5, and 17E6, while 21H6 and 14E2 precipitate a band at ~200 kDa. 14D9 precipitates both patterns. M21 melanoma (B), M21-L αv-deficient melanoma (C), and UCLA-P3 cells (D) were cell-surface labelled and immunoprecipitated. Precipitation was with antibodies LM142 (anti-αv: lane a), LM609 (anti-αvβ3: lane b), 17E6 (lane c), 20A9 (lane d), AP3 (anti-β3: lane e), P4C10 (anti-β1: lane f), and B5-1A9 (anti-β5: lane g). Positions of molecular weight markers are shown as small arrows to left of the panel, with weights in kDa (shown in C). The position of αv (open arrow), β1, β5, β3, (closed arrows) and ~200 kDa chains is indicated to the right of each panel. Note the β3 chain precipitated from M21-L cells (lane e) permeabilized during surface labelling.

Mitjans, F., Sander, D., Adán, J., Sutter, A., Martinez, J. M., Jaggle, C. S., ... & Goodman, S. L. (1995). An anti-alpha v-integrin antibody that blocks integrin function inhibits the development of a human melanoma in nude mice. Journal of cell science, 108(8), 2825-2838.

Figure 2 Reaction pattern of monoclonal antibodies in ELISAs and CELISAs

Figure 2 Reaction pattern of monoclonal antibodies in ELISAs and CELISAs

Antibodies were compared in ELISA and cell ELISA (CELISA). Reagents and specificities are discussed fully in text. +, antibodies bind; −, antibodies do not bind. The antibodies characterized in this study are in bold, standard reagent antibodies are italicised. Immunogen: αvβ3 = immobilized human placental integrin αvβ3. M21= viable M21 cells. Purified αvβ3 and αIIbβ3 were immobilized on 96-well plates for ELISA. The cell lines M21, M21-L and M21-L-IIb, and UCLA-P3 were grown and fixed on the plates and tested for antibody reactivity in CELISA. Specificity: (See text) αv = αv chain of mammalian integrins. 200 kDa = unknown 200,000 Da surface protein of melanoma cells. αvβ5 = integrin αvβ5. β1 = integrin β1 chain. β3 = integrin β3 chain.

Mitjans, F., Sander, D., Adán, J., Sutter, A., Martinez, J. M., Jaggle, C. S., ... & Goodman, S. L. (1995). An anti-alpha v-integrin antibody that blocks integrin function inhibits the development of a human melanoma in nude mice. Journal of cell science, 108(8), 2825-2838.

Figure 3 FACS analysis of the alpha-v group antibody binding on M21 and M21-L human melanoma cells.

Figure 3 FACS analysis of the alpha-v group antibody binding on M21 and M21-L human melanoma cells.

Cells were indirectly labelled with antibodies. Propidium iodide was added and 15,000 cells per sample were acquired and analysed after gating. The open peak represents the binding of the fluorescently-labelled second layer antibody alone. The closed peak, the intensity of the primary and secondary together. Vertical axis shows cell number, horizontal axis shows log fluorescence intensity. M21 express surface αv integrin, whereas M21-L does not. The pattern of staining for the alpha-v group antibodies (17E6, 20A9, 23G5, 14D9.F8) closely matches the LM142 (αv-specific) and LM609 (αvβ3- specific) staining. They bind M21 but not M21-L cells. Antibody 9.2.27 reacts with a melanoma proteoglycan. 14E2 and 21H6 recognise a 200 kDa melanoma surface protein. Their staining patterns are discrete from those of the alpha-v group and similar for both cell lines.

Mitjans, F., Sander, D., Adán, J., Sutter, A., Martinez, J. M., Jaggle, C. S., ... & Goodman, S. L. (1995). An anti-alpha v-integrin antibody that blocks integrin function inhibits the development of a human melanoma in nude mice. Journal of cell science, 108(8), 2825-2838.

Figure 4 17E6 perturbs M21 adhesion to vitronectin but not to other matrix components.

Figure 4 17E6 perturbs M21 adhesion to vitronectin but not to other matrix components.

The inhibitory effect of 17E6 (αv, closed circles) and P4C10 (β1, open circles) on M21 cell attachment to vitronectin (A), laminin (B) or collagen type I (C) was studied. 17E6 only blocked cell attachment on vitronectin-coated surfaces. Whilst P4C10 only blocked cell attachment on collagen type I and laminin.

Mitjans, F., Sander, D., Adán, J., Sutter, A., Martinez, J. M., Jaggle, C. S., ... & Goodman, S. L. (1995). An anti-alpha v-integrin antibody that blocks integrin function inhibits the development of a human melanoma in nude mice. Journal of cell science, 108(8), 2825-2838.

Figure 5 Species-dependent inhibition of cell attachment by 17E6 and species-independent inhibition by cilengitide.

Figure 5 Species-dependent inhibition of cell attachment by 17E6 and species-independent inhibition by cilengitide.

Representative cell adhesion inhibition assay comparing human melanoma cell line WM164 (circles), monkey cell line CV-1 (triangles), and rat cell line Rat-1 (boxes). Filled symbols show response to 17E6, and open symbols show response to the RGD-peptide cilengitide. Human and monkey cells are inhibited, and rat cells are not affected by 17E6. Each species is inhibited by cilengitide.

Mahalingam, B., Van Agthoven, J. F., Xiong, J. P., Alonso, J. L., Adair, B. D., Rui, X., ... & Goodman, S. L. (2014). Atomic basis for the species-specific inhibition of αV integrins by mAb 17E6 is revealed by the crystal structure of αVβ3 ectodomain-17E6 Fab complex. Journal of Biological Chemistry, jbc-M113.

Figure 6 Effect of αv-integrin antagonists on the distribution pattern of αv-integrins and actin cytoskeleton.

Figure 6 Effect of αv-integrin antagonists on the distribution pattern of αv-integrins and actin cytoskeleton.

M21 cells were seeded and cultured as described in Materials and methods. After αv-integrin antagonist treatments cells were fixed and processed for immunofluorescence analysis. Cells treated with mAbs were only incubated with a FITC-conjugated anti-mouse immunoglobulin but control cells and cells treated with cRGD and cRAD were first incubated with mAbs 17E6 followed by incubation with the secondary FITC-conjugated anti-mouse immunoglobulin. F-actin was detedcted using TRITC-conjugated phalloidin. Three-dimensional projection of 9 (0 min) or 16 (30 min) dual colour confocal x, y sections is shown. Z-step size between xy sections was 0.3 mm. Cells treated with antagonists became rounded, losing their flattened form, then they enlarged the xz distance and they needed more sections (16 instead of 9) to maintain the z-step for series. Calibration: pixel size (x, y) is 0.31 μm/pixel, z-step is 0.3 μm. One representative experiment of three is shown. Scale bar 10 μm.

Castel, S., Pagan, R., García, R., Casaroli-Marano, R. P., Reina, M., Mitjans, F., ... & Vilaró, S. (2000). Alpha v integrin antagonists induce the disassembly of focal contacts in melanoma cells. European journal of cell biology, 79(7), 502-512.


Specifications

  • Host Species
  • Mouse
  • Type
  • Mouse IgG
  • Specificity
  • Human αV Integrins
  • Species Reactivity
  • Human
  • Clone
  • 17E6
  • Applications
  • ELISA, FuncS
  • Conjugate
  • Unconjugated

Product Property

  • Purification
  • Protein A Purified
  • Format
  • Liquid
  • Concentration
  • Batch Specific
  • Buffer
  • PBS
  • Storage
  • Store at 4°C for short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Applications

  • Application Notes
  • This antibody has been tested for use in Enzyme-linked Immunosorbent Assay, Functional Assay.

Target

  • Alternative Names
  • ITGAV; integrin, alpha V; CD51; MSK8; VNRA; VTNR; integrin alpha-V; integrin alphaVbeta3; vitronectin receptor subunit alpha; antigen identified by monoclonal antibody L230; integrin, alpha V (vitronectin receptor, alpha polypeptide, antigen CD51); αVβ1; αVβ3; αVβ5; αVβ6; αVβ8

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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Fab Fragment Antibody

CAT Product Name Application Type
PABX-191-F(E) Mouse Anti-αV Integrins Recombinant Antibody (clone 17E6); Fab Fragment ELISA, FuncS Mouse Fab

scFv Fragment Antibody

CAT Product Name Application Type
PABX-191-S(P) Mouse Anti-αV Integrins Recombinant Antibody (clone 17E6); scFv Fragment ELISA, FuncS Mouse scFv

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For Research Use Only. Not For Clinical Use.

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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