Human Anti-ITGAL Recombinant Antibody (clone AL-57) (CAT#: PABW-159)

Recombinant Human Antibody (AL-57) is capable of binding to ITGAL, expressed in HEK 293 cells. Expressed as the combination of a heavy chain (HC) containing VH from anti-ITGAL mAb and CH1-3 region of human IgG and a light chain (LC) encoding VL from anti-ITGAL proteins mAb and CL of human light chain. Exists as a disulfide linked dimer of the HC and LC hetero-dimer under non-reducing condition. AL-57 functions as an ICAM-1 mimetic antibody that exhibits several key features of the ICAM-1/LFA-1 interaction. Not only do AL-57 and ICAM-1 both bind progressively better as LFA-1 affinity increases, they both require Mg2+ for binding.


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SPR

Figure 1 SPR analysis of binding by the αL I domains to Abs AL-57 and MHM24.

Figure 1 SPR analysis of binding by the αL I domains to Abs AL-57 and MHM24.

The HA (K287C/K294C), IA (L161C/F299C), or low-affinity WT I domain was perfused onto immobilized Abs in the presence of 1 mM MgCl2. The concentration series for MHM24 was 3.91, 7.81, 15.63, 31.25, 62.5, 125, 250, and 500 nM. For AL-57, the concentration series was 15.6, 31.3, 62.5, 125, and 250 nM for the HA I domain and 31.25, 62.5, 125, 250, and 500 nM for WT and IA I domains. In all cases, higher concentrations gave higher responses (except that differences are not visible for WT with AL-57 in Mg2+).

Shimaoka, M., Kim, M., Cohen, E. H., Yang, W., Astrof, N., Peer, D., ... & Springer, T. A. (2006). AL-57, a ligand-mimetic antibody to integrin LFA-1, reveals chemokine-induced affinity up-regulation in lymphocytes. Proceedings of the National Academy of Sciences, 103(38), 13991-13996.

SPR

Figure 2 SPR analysis of binding by the αL I domains to Abs AL-57 and MHM24.

Figure 2 SPR analysis of binding by the αL I domains to Abs AL-57 and MHM24.

The HA (K287C/K294C), IA (L161C/F299C), or low-affinity WT I domain was perfused onto immobilized Abs in the presence of 10 mM EDTA. The concentration series for MHM24 was 3.91, 7.81, 15.63, 31.25, 62.5, 125, 250, and 500 nM. For AL-57, the concentration series was 15.6, 31.3, 62.5, 125, and 250 nM for the HA I domain and 31.25, 62.5, 125, 250, and 500 nM for WT and IA I domains. In all cases, higher concentrations gave higher responses (except that differences are not visible for HA with AL-57 in EDTA).

Shimaoka, M., Kim, M., Cohen, E. H., Yang, W., Astrof, N., Peer, D., ... & Springer, T. A. (2006). AL-57, a ligand-mimetic antibody to integrin LFA-1, reveals chemokine-induced affinity up-regulation in lymphocytes. Proceedings of the National Academy of Sciences, 103(38), 13991-13996.

FC

Figure 3 Acidic residue D101 of AL-57 is critical for binding.

Figure 3 Acidic residue D101 of AL-57 is critical for binding.

Binding of parental and mutant AL-57 IgG (3 μg/ml) to the isolated, locked HA I domain expressed on K562 cells was examined by immunofluorescent flow cytometry in the presence of 1 mM Mg2+ (filled histograms) or 5 mM EDTA (open histograms). Binding of AL-57D101A in Mg2+ and EDTA is hardly distinguishable from background binding of control IgG.

Shimaoka, M., Kim, M., Cohen, E. H., Yang, W., Astrof, N., Peer, D., ... & Springer, T. A. (2006). AL-57, a ligand-mimetic antibody to integrin LFA-1, reveals chemokine-induced affinity up-regulation in lymphocytes. Proceedings of the National Academy of Sciences, 103(38), 13991-13996.

FC

Figure 4 Binding of AL-57 IgG to WT LFA-1 activated by agonists as determined by immunofluorescent flow cytometry.

Figure 4 Binding of AL-57 IgG to WT LFA-1 activated by agonists as determined by immunofluorescent flow cytometry.

K562 transfectants expressing WT LFA-1 and IL-15-cultured human primary T lymphocytes were stained for 20 min at 37°C with 20 μg/ml AL-57, TS2/4, or isotype-matched human or mouse control IgG in Hepes-buffered saline containing 1 mM MgCl2/1 mM CaCl2, 5 mM MgCl2/1 mM EGTA, or 1 mM MnCl2. Cells were washed and stained with FITC-conjugated goat anti-human or anti-mouse Abs. Staining with AL-57 and TS2/4 is shown as solid lines, and background staining with control IgGs is shown as dashed lines. Numbers within the panels show the specific mean fluorescence intensity of AL-57 and TS2/4.

Shimaoka, M., Kim, M., Cohen, E. H., Yang, W., Astrof, N., Peer, D., ... & Springer, T. A. (2006). AL-57, a ligand-mimetic antibody to integrin LFA-1, reveals chemokine-induced affinity up-regulation in lymphocytes. Proceedings of the National Academy of Sciences, 103(38), 13991-13996.

FC

Figure 5 The kinetics of LFA-1 activation after stimulation with CXCL-12.

Figure 5 The kinetics of LFA-1 activation after stimulation with CXCL-12.

Representative histograms for a time course of AL-57 epitope expression are shown. T lymphocytes were stimulated with 50 ng/ml CXCL-12 for the indicated time period. Cells were stained with Fab AL-57 or control Fab labeled with Alexa 488 for 2 min by adding Abs 2 min before the end of stimulation. Cells were then fixed and subjected to flow cytometry. Background staining with control Fab is shown in the open histograms. Numbers within the panels show specific mean fluorescence intensity values for Fab AL-57. At 0, 10, and 20 min, differences between AL-57 Fab and control Fab binding are hardly visible.

Shimaoka, M., Kim, M., Cohen, E. H., Yang, W., Astrof, N., Peer, D., ... & Springer, T. A. (2006). AL-57, a ligand-mimetic antibody to integrin LFA-1, reveals chemokine-induced affinity up-regulation in lymphocytes. Proceedings of the National Academy of Sciences, 103(38), 13991-13996.


Specifications

  • Immunogen
  • Human integrin, alpha L (antigen CD11A (p180), lymphocyte function-associated antigen 1; alpha polypeptide)
  • Host Species
  • Human
  • Derivation
  • Human
  • Type
  • Human IgG
  • Specificity
  • Human ITGAL
  • Species Reactivity
  • Human
  • Clone
  • AL-57
  • Applications
  • WB

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
  • ITGAL; integrin, alpha L (antigen CD11A (p180), lymphocyte function-associated antigen 1; alpha polypeptide); CD11A; LFA-1; LFA1A; integrin alpha-L; LFA-1A; LFA-1 alpha; integrin gene promoter; CD11 antigen-like family member A; lymphocyte function-associated antigen 1; leukocyte adhesion glycoprotein LFA-1 alpha chain; leukocyte function-associated molecule 1 alpha chain; antigen CD11A (p180), lymphocyte function-associated antigen 1, alpha polypeptide

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.

Downloads

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Humanized Antibody

CAT Product Name Application Type
TAB-024 Anti-Human CD11A Recombinant Antibody (Efalizumab) Neut, ELISA, IF, IP, FuncS, FC, WB IgG1 - kappa

Chimeric Antibody

CAT Product Name Application Type
TAB-755LC Anti-human ITGAL Recombinant Antibody (Chimeric S6F1) (TAB-755LC) ELISA Chimeric antibody (mouse/human)
TAB-755LC-S(P) Anti-human ITGAL scFv Fragment (Chimeric S6F1) ELISA Chimeric antibody (mouse/human)
TAB-755LC-F(E) Anti-human ITGAL Fab Fragment (Chimeric S6F1) ELISA Chimeric antibody (mouse/human)

Neutralizing Antibody

CAT Product Name Application Type
NEUT-1559CQ Recombinant Mouse Anti-ITGAL Antibody (38) FC, ICC/IF, IHC-Fr, IP, Neut IgG2a
NEUT-1563CQ Recombinant Mouse Anti-ITGAL Antibody (CBL227) FC, IHC, IP, Neut IgG2a
NEUT-1565CQ Recombinant Rat Anti-Itgal Antibody (121/7) FC, IP, Neut, WB IgG2a
NEUT-1568CQ Recombinant Rat Anti-Itgal Antibody (I21/7) Neut, FC, IHC, IHC-Fr, IP, ICC IgG2a, κ

Rabbit Monoclonal Antibody

CAT Product Name Application Type
MOR-1878 Hi-Affi™ Recombinant Rabbit Anti-ITGAL Monoclonal Antibody (DS1878AB) BL, FC, ICC/IF, IHC-P, IP, WB IgG

Neuroscience Antibody

CAT Product Name Application Type
NS-018CN Human Anti-ITGAL Recombinant Antibody (clone MHM24) FC, ELISA, Inhib, FuncS Human IgG1

ADCC Enhanced Antibody

CAT Product Name Application Type
AFC-TAB-108 Afuco™ Anti-ITGAL ADCC Recombinant Antibody (Odulimomab), ADCC Enhanced Neut, ELISA, IF, IP, FuncS, FC ADCC enhanced antibody
AFC-TAB-024 Afuco™ Anti-ITGAL ADCC Recombinant Antibody (Efalizumab), ADCC Enhanced Neut, ELISA, IF, IP, FuncS, FC ADCC enhanced antibody

Single-domain Antibody

CAT Product Name Application Type
MOB-0066SW Recombinant Llama Anti-ITGAL Single Domain Antibody (138-B10) ELISA, WB Llama VHH

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