Recombinant Anti-HLA-DR antibody [TAL 1B5] (ab20181)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [TAL 1B5] to HLA-DR
- Suitable for: ICC/IF, WB, IHC-P, Flow Cyt
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-HLA-DR antibody [TAL 1B5]
See all HLA-DR primary antibodies -
Description
Mouse monoclonal [TAL 1B5] to HLA-DR -
Host species
Mouse -
Tested applications
Suitable for: ICC/IF, WB, IHC-P, Flow Cytmore details -
Species reactivity
Reacts with: Human -
Immunogen
Full length native protein (purified). This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Raji and Daudi whole cell lysates; Human colon lysate. IHC-P: Human skin and tonsil tissue sections. Flow Cyt: Human peripheral blood mononuclear cells (PBMCs). ICC/IF: Raji cells.
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General notes
This product has switched from a hybridoma to recombinant production method on 23 September 2022.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
TAL 1B5 -
Myeloma
P3-NS1/1-Ag4-1 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab20181 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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ICC/IF |
1/1000.
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WB | (1) |
Use a concentration of 1 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 29 kDa).
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IHC-P | (3) |
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt |
Use a concentration of 0.008 µg/ml.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Notes |
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ICC/IF
1/1000. |
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 29 kDa). |
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt
Use a concentration of 0.008 µg/ml. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Target
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Function
Binds 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 similarities
Belongs to the MHC class II family.
Contains 1 Ig-like C1-type (immunoglobulin-like) domain. -
Post-translational
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 localization
Cell 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
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Database links
- Entrez Gene: 3122 Human
- Omim: 142860 Human
- SwissProt: P01903 Human
- Unigene: 520048 Human
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Alternative names
- DASS-397D15.1 antibody
- DR alpha chain antibody
- DR alpha chain precursor antibody
see all
Images
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ab20181 staining HLA DR in Raji cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab20181 at 1/1000 dilution and ab7291, Mouse monoclonal to alpha Tubulin at 1/1000 dilution. Cells were then incubated with ab150119, Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed, at 1/1000 dilution (shown in red) and ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488), preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). -
All lanes : Anti-HLA-DR antibody [TAL 1B5] (ab20181) at 1/1000 dilution
Lane 1 : Raji whole cell lysate
Lane 2 : Daudi whole cell lysate
Lane 3 : HEK-293 whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 29 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab20181 observed at 35 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.
ab20181 was shown to react with HLA-DR in Western blot. Membranes were blocked with 3% milk in TBS-T (0.1% Tween®) before incubation with ab20181 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4°C at 1 µg/ml and a 1:20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1:20000 dilution for 1 h at room temperature before imaging.
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Immunohistochemical analysis of paraffin-embedded human skin tissue labeling HLA-DR with ab20181 at 0.1 µg/ml followed by Leica DS9800 (Bond™ Polymer Refine Detection). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab20181, 0.1ug/ml, for 15 mins at room temperature and was then detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling HLA-DR with ab20181 at 0.1 µg/ml followed by Leica DS9800 (Bond™ Polymer Refine Detection). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab20181, 0.1ug/ml, for 15 mins at room temperature and was then detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
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Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab20181 (right) or mouse IgG1κ (ab170190) isotype (left). PBMCs were incubated for 30 min on ice in 1x PBS containing 10μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by followed by staining with CD19 APC. PBMCs were then fixed in 4.2% formaldehyde and permeabilised in 0.1% saponin before staining with the antibody (ab20181) or mouse IgG1κ (ab170190) isotype (1x106 in 100μl; at 0.008μg/ml) for 30 min on ice. The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor ® 488, pre-adsorbed) (ab150117) was used at 1:2000 dilution for 30 min on ice. Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on alive lymphocytes.
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Anti-HLA-DR antibody [TAL 1B5] (ab20181) at 0.667 µg/ml + whole tissue lysate prepared from human colon at 50 µg
Secondary
HRP conjugated goat anti-mouse polyclonal at 1/3000 dilution
Developed using the ECL technique.
Predicted band size: 29 kDa
Observed band size: 33 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsThis image was generated from a previous batch made using the hybridoma production method.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (40)
ab20181 has been referenced in 40 publications.
- Lim CY et al. CD40 Agonists Alter the Pancreatic Cancer Microenvironment by Shifting the Macrophage Phenotype toward M1 and Suppress Human Pancreatic Cancer in Organotypic Slice Cultures. Gut Liver 16:645-659 (2022). PubMed: 34933280
- Duan J et al. Multidimensional profiling depicts infiltrating immune cell heterogeneity in the tumor microenvironment of stage IA non-small cell lung cancer. Thorac Cancer 13:947-955 (2022). PubMed: 35150094
- Thirimanne HN et al. Global and context-specific transcriptional consequences of oncogenic Fbw7 mutations. Elife 11:N/A (2022). PubMed: 35225231
- Kaneko N et al. Temporal changes in T cell subsets and expansion of cytotoxic CD4+ T cells in the lungs in severe COVID-19. Clin Immunol 237:108991 (2022). PubMed: 35364330
- Lee JS et al. Expression of Inflammatory Markers in the Muscles of Patients with Idiopathic Inflammatory Myopathy According to the Presence of Interstitial Lung Disease. J Clin Med 11:N/A (2022). PubMed: 35683408