Clone:
REA527
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Alternative names:
H2Kd, H2Dd,
H-2K
d
,
H-2D
d

Extended validation for H-2Kd/H-2Dd Antibody, anti-mouse, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA527
34-1-2S++
34-5-8S-
REAL411++
Cells were incubated with an excess of purified unconjugated H-2Kd/H-2Dd (REA527) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for H-2Kd/H-2Dd. C57BL6/6 splenocytes were stained with H-2Kd/H-2Dd antibodies and with a suitable counterstaining. As a control H-2Kd/H-2Dd antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for H-2Kd/H-2Dd. C57BL6/6 splenocytes were stained with H-2Kd/H-2Dd antibodies and with a suitable counterstaining. As a control H-2Kd/H-2Dd antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for H-2Kd/H-2Dd. C57BL6/6 splenocytes were stained with H-2Kd/H-2Dd antibodies and with a suitable counterstaining. As a control H-2Kd/H-2Dd antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for H-2Kd/H-2Dd. C57BL6/6 splenocytes were stained with H-2Kd/H-2Dd antibodies and with a suitable counterstaining. As a control H-2Kd/H-2Dd antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using H-2Kd/H-2Dd (REA527). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using H-2Kd/H-2Dd (REA527). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using H-2Kd/H-2Dd (REA527). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for H-2Kd/H-2Dd Antibody, anti-mouse, REAfinity™

Overview

Clone REA527 recognizes the mouse H-2K
d
and H-2D
d
MHC class I alloantigens expressed on mouse MHC class I–positive cells of the H-2K
d
/H-2D
d
haplotype. It also cross-reacts with the H-2Kb, s, r, q, or p haplotypes.
Additional information: Clone REA527 displays negligible binding to Fc receptors.

Alternative names

H2Kd, H2Dd, H-2Kd, H-2Dd

Detailed product information

Technical specifications

CloneREA527
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenH-2Kd/H-2Dd
Alternative names of antigenH2Kd, H2Dd,
H-2K
d
,
H-2D
d
Molecular mass of antigen [kDa]39(average of subunits)
Distribution of antigenother
Entrez Gene ID14964, 14972
RRIDAB_2877029, AB_2651957, AB_2651958, AB_2651961, AB_2651962, AB_2651965, AB_2651966, AB_2651967, AB_2651968, AB_2651969, AB_2651970, AB_2651971, AB_2651972

Resources for H-2Kd/H-2Dd Antibody, anti-mouse, REAfinity™

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

References for H-2Kd/H-2Dd Antibody, anti-mouse, REAfinity™

Publications

  1. Silver, E. T. et al. (2000) Ly-49P activates NK-mediated lysis by recognizing H-2Dd. J. Immunol. 165(4): 1771-1781
  2. Schaffar-Deshayes, L. et al. (1983) Detection of H-2Dd and H-2Kd molecules in purified Rauscher leukemia virus. Int. J. Cancer 31(3): 329-335
  3. Mitaksov, V. et al. (2006) Structural definition of the H-2Kd peptide-binding motif. J. Biol. Chem. 281(15): 10618-10625

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