Human Anti-HLA-A Recombinant Antibody (clone HYB3) (CAT#: PABC-072)

Recombinant Human Antibody (HYB3) is capable of binding to HLA-A1. MAGE-A1, expressed in HEK 293 cells. Expressed as the combination of a heavy chain (HC) containing VH from anti-HLA-A1. MAGE-A1 mAb and CH1-3 region of human IgG1 and a light chain (LC) encoding VL from anti-HLA-A1 mAb and CL of human kappa light chain. Exists as a disulfide linked dimer of the HC and LC hetero-dimer under non-reducing condition. It recognizes the C-terminal half of the MAGE-A1 peptide, the HLA-A1 alpha1-helix, and N-terminal residues of the alpha2-helix, accompanied by a large tilting angle between the two types of molecules within the complex.

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ELISA

Figure 1 Fine specificity of the affinity-matured clones by phage-ELISA.

pMHC complexes were refolded using each peptide and coated via streptavidin on an ELISA plate. Equal concentrations of phage-Abs were incubated for 1 h at room temperature. After extensive washing, bound phages were detected with an anti-M13 mAb coupled to HRP.

Chames, P., Willemsen, R. A., Rojas, G., Dieckmann, D., Rem, L., Schuler, G., ... & Hoogenboom, H. R. (2002). TCR-like human antibodies expressed on human CTLs mediate antibody affinity-dependent cytolytic activity. The Journal of Immunology, 169(2), 1110-1118.

FC

Figure 2 scFv G8 and Hyb3 specifically recognize M1/A1 on cells.

Flow cytometry of HLA-A1+ B cells (APD) with or without MAGE A1 peptide with Hyb3. Histograms showing binding of scFv G8 and Hyb3 at a concentration of 10 µg/mL to APD cells pulsed with peptide MAGE A1. Phage Hyb3 (purple) were used as a positive control for binding to APD cells.

Saeed, M., van Brakel, M., Zalba, S., Schooten, E., Rens, J. A., Koning, G. A., ... & ten Hagen, T. L. (2016). Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody. International journal of nanomedicine, 11, 955.

IF

Figure 3 Binding of immunoliposomes to APD cells.

Confocal images of APD cells (500,000 cells per mL) pulsed with peptide and incubated with immunoliposomes for 2 hours at 37°C.

Saeed, M., van Brakel, M., Zalba, S., Schooten, E., Rens, J. A., Koning, G. A., ... & ten Hagen, T. L. (2016). Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody. International journal of nanomedicine, 11, 955.

WB

Figure 4 Purification of scFv Hyb3 proteins by IMAC.

Western blot analyses of purity and size of purified scFv proteins. Fractions 7 and 8 consist of the peak fractions from the chromatogram and were run on SDS-PAGE and subjected by Western blot using anti-HIS mAb HRP.

Saeed, M., van Brakel, M., Zalba, S., Schooten, E., Rens, J. A., Koning, G. A., ... & ten Hagen, T. L. (2016). Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody. International journal of nanomedicine, 11, 955.


Specifications

  • Immunogen
  • Human major histocompatibility complex, class I, A
  • Host Species
  • Human
  • Type
  • Human IgG
  • Specificity
  • Human HLA-A
  • Species Reactivity
  • Human
  • Clone
  • HYB3
  • Applications
  • WB, ELISA, IF, FC

Applications

  • Application Notes
  • This antibody has been reported for use in Western Blot, Enzyme-Linked Immunosorbent Assay, Immunofluorescence and Flow Cytometry.

Target

  • Alternative Names
  • HLA-A; major histocompatibility complex, class I, A; HLAA; HLA class I histocompatibility antigen, A-1 alpha chain; antigen presenting molecule; leukocyte antigen class I-A; MHC class I antigen HLA-A heavy chain

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.

See other products for "HLA-A"


For research use only. Not intended for any clinical use.

* 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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