Clone:
REA1133
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
CR1, C3BR, C4BR, KN, C3b/C4b-R

Extended validation for CD35 Antibody, anti-human, 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 REA1133
E11++
Cells were incubated with an excess of purified unconjugated CD35 (REA1133) 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 CD35. Human peripheral blood mononuclear cells (PBMCs) were stained with CD35 antibodies and with a suitable counterstaining. As a control, CD35 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 CD35. Human peripheral blood mononuclear cells (PBMCs) were stained with CD35 antibodies and with a suitable counterstaining. As a control, CD35 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 CD35. Human peripheral blood mononuclear cells (PBMCs) were stained with CD35 antibodies and with a suitable counterstaining. As a control, CD35 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 CD35. Human peripheral blood mononuclear cells (PBMCs) were stained with CD35 antibodies and with a suitable counterstaining. As a control, CD35 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 CD35 (REA1133). 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 CD35 (REA1133). 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 CD35 (REA1133). 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 CD35 Antibody, anti-human, REAfinity™

Overview

Clone REA1133 recognizes the human cell surface protein CD35, the C3b/C4b receptor, also known as CR1 or complement receptor type I. Four CD35 allotypes with different molecular weights are known; 190, 220, 250, and 280 kDa. CD35 is expressed on hematopoietic cells such as granulocytes, monocytes, B cells, subsets of NK and T cells, erythrocytes, and follicular dendritic cells (DCs). Known functions of CD35 include transport of C3 complexes on erythrocytes, complement activation, phagocytosis, and inhibition of the activation-induced proliferation of T and B cells.
Additional information: Clone REA1133 displays negligible binding to Fc receptors.

Alternative names

CR1, C3BR, C4BR, KN, C3b/C4b-R

Detailed product information

Technical specifications

CloneREA1133
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, non-human primate
Cross-reactivity
rhesus monkey (
Macaca mulatta
)
,
cynomolgus monkey (
Macaca fascicularis
)
,
chimpanzee (
Pan troglodytes
)
,
olive baboon (
Papio anubis
)
AntigenCD35
Alternative names of antigenCR1, C3BR, C4BR, KN, C3b/C4b-R
Molecular mass of antigen [kDa]220
Distribution of antigendendritic cells, T cells, NK cells
Entrez Gene ID1378
RRIDAB_2751710, AB_2751766, AB_2751711, AB_2751767, AB_2751712, AB_2751768, AB_2751713, AB_2751769, AB_2751714, AB_2751770, AB_2751715, AB_2921919, AB_2921888, AB_2751765

Resources for CD35 Antibody, anti-human, REAfinity™

Certificates

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

References for CD35 Antibody, anti-human, REAfinity™

Publications

  1. Reynes, M. et al. (1985) Human follicular dendritic cells express CR1, CR2, and CR3 complement receptor antigens. J Immunol 135: 2687-2694
  2. Klickstein, L. B. et al. (1988) Identification of distinct C3b and C4b recognition sites in the human C3b/C4b receptor (CR1, CD35) by deletion mutagenesis. J. Exp. Med. 168: 1699-1717
  3. Reilly, B. et al. (2010)
    Human peripheral CD4
    +
    T-cells expressing complement receptor 1 (CR1, CD35).
    J Immunol 184(1 Supplement): 138.3

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