Key features and details
- One-wash 90 minute protocol
- Sensitivity: 5 pg/ml
- Range: 31.3 pg/ml - 2000 pg/ml
- Sample type: Cell culture extracts, Tissue Homogenate
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Product nameHuman HLA-DR ELISA Kit
See all HLA-DR kits
Intra-assay Sample n Mean SD CV% Cell extract 5 3.6% Inter-assay Sample n Mean SD CV% Cell extract 3 12.7%
Sample typeCell culture extracts, Tissue Homogenate
Assay typeSandwich (quantitative)
Range31.3 pg/ml - 2000 pg/ml
Sample specific recovery Sample type Average % Range Cell culture extracts 98 89% - 105% Tissue Homogenate 94 86% - 100%
Assay time1h 30m
Assay durationOne step assay
Species reactivityReacts with: Human
Human HLA-DR ELISA Kit (ab223593) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of HLA-DR protein in tissue homogenate and cell culture extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human HLA-DR with 5 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
HLA class II histocompatibility antigen, DR alpha chain (HLA-DR) is the alpha chain of a cell surface receptor heterodimer that binds peptides for presentation to CD4 T cells. The alpha chain is fairly constant between individuals and pairs with varying alleles of the beta chain. HLA-DR has been implicated in multiple auto-immune conditions, as well as graft vs host disease.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
PlatformPre-coated microplate (12 x 8 well strips)
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 10X Human HLA-DR Capture Antibody 1 x 600µl 10X Human HLA-DR Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 4BI 1 x 6ml Human HLA-DR Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml
FunctionBinds peptides derived from antigens that access the endocytic route of antigen presenting cells (APC) and presents them on the cell surface for recognition by the CD4 T-cells. The peptide binding cleft accommodates peptides of 10-30 residues. The peptides presented by MHC class II molecules are generated mostly by degradation of proteins that access the endocytic route, where they are processed by lysosomal proteases and other hydrolases. Exogenous antigens that have been endocytosed by the APC are thus readily available for presentation via MHC II molecules, and for this reason this antigen presentation pathway is usually referred to as exogenous. As membrane proteins on their way to degradation in lysosomes as part of their normal turn-over are also contained in the endosomal/lysosomal compartments, exogenous antigens must compete with those derived from endogenous components. Autophagy is also a source of endogenous peptides, autophagosomes constitutively fuse with MHC class II loading compartments. In addition to APCs, other cells of the gastrointestinal tract, such as epithelial cells, express MHC class II molecules and CD74 and act as APCs, which is an unusual trait of the GI tract. To produce a MHC class II molecule that presents an antigen, three MHC class II molecules (heterodimers of an alpha and a beta chain) associate with a CD74 trimer in the ER to form an heterononamer. Soon after the entry of this complex into the endosomal/lysosomal system where antigen processing occurs, CD74 undergoes a sequential degradation by various proteases, including CTSS and CTSL, leaving a small fragment termed CLIP (class-II-associated invariant chain peptide). The removal of CLIP is facilitated by HLA-DM via direct binding to the alpha-beta-CLIP complex so that CLIP is released. HLA-DM stabilizes MHC class II molecules until primary high affinity antigenic peptides are bound. The MHC II molecule bound to a peptide is then transported to the cell membrane surface. In B-cells, the interaction between HLA-DM and MHC class II molecules is regulated by HLA-DO. Primary dendritic cells (DCs) also to express HLA-DO. Lysosomal miroenvironment has been implicated in the regulation of antigen loading into MHC II molecules, increased acidification produces increased proteolysis and efficient peptide loading.
Sequence similaritiesBelongs to the MHC class II family.
Contains 1 Ig-like C1-type (immunoglobulin-like) domain.
modificationsUbiquitinated by MARCH1 or MARCH8 at Lys-244 leading to down-regulation of MHC class II. When associated with ubiquitination of the beta subunit of HLA-DR: HLA-DRB4 'Lys-254', the down-regulation of MHC class II may be highly effective.
Cellular localizationCell membrane. Endoplasmic reticulum membrane. Golgi apparatus > trans-Golgi network membrane. Endosome membrane. Lysosome membrane. Late endosome membrane. The MHC class II complex transits through a number of intracellular compartments in the endocytic pathway until it reaches the cell membrane for antigen presentation.
- Information by UniProt
- DR alpha chain
- DR alpha chain precursor
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
Background-subtracted data values (mean +/- SD) are graphed.
The concentrations of HLA-DR were measured in duplicates, interpolated from the HLA-DR standard curves corrected for sample dilution and extract load. Undiluted samples are as follows: Daudi cell extract 32.3 µg/mL, Raji cell extract 490 µg/mL, and human skin extract 500 µg/mL. The interpolated dilution factor and corrected values are plotted (mean +/- SD, n=2). The mean HLA-DR concentration was determined to be 52.0 pg/µg in Daudi Extract, 1.93 pg/µg in Raji Extract and 0.85 pg/µg in human skin homogenate extract.
ab223593 has not yet been referenced specifically in any publications.