Recombinant Anti-HLA-DR antibody [TAL 1B5] - BSA and Azide free (ab176408)
![Western blot - Anti-HLA-DR antibody [TAL 1B5] - BSA and Azide free (ab176408)](https://www.abcam.com/ps/products/176/ab176408/Images/ab176408-491765-anti-hla-dr-antibody-tal-1b5-western-blot-raji-daudi-hek-293.jpg "Western blot - Anti-HLA-DR antibody [TAL 1B5] - BSA and Azide free (ab176408)")
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 - BSA and Azide free
- Suitable for: 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] - BSA and Azide free
See all HLA-DR primary antibodies -
Description
Mouse monoclonal [TAL 1B5] to HLA-DR - BSA and Azide free -
Host species
Mouse -
Tested applications
Suitable for: WB, IHC-P, Flow Cytmore details -
Species reactivity
Reacts with: Human -
Immunogen
Tissue, cells or virus corresponding to HLA-DR. Bristol 8 separated alpha chain preparation
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Positive control
- WB: Raji and Daudi whole cell lysates. IHC-P: Human skin and tonsil tissue sections. Flow Cyt: Human peripheral blood mononuclear cells (PBMCs).
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General notes
ab176408 is the carrier-free version of ab20181.
This product has switched from a hybridoma to recombinant production method on 23 September 2022.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading... -
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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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab176408 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 |
|---|---|---|
| WB |
Use at an assay dependent concentration. Detects a band of approximately 29, 35 kDa (predicted molecular weight: 29 kDa).
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| IHC-P | (2) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| Flow Cyt |
Use at an assay dependent concentration.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
| Notes |
|---|
|
WB
Use at an assay dependent concentration. Detects a band of approximately 29, 35 kDa (predicted molecular weight: 29 kDa). |
|
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
|
Flow Cyt
Use at an assay dependent concentration. 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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Western blot - Anti-HLA-DR antibody [TAL 1B5] - BSA and Azide free (ab176408)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
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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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [TAL 1B5] - BSA and Azide free (ab176408)](https://www.abcam.com/ps/products/176/ab176408/Images/ab176408-491767-anti-hla-dr-antibody-tal-1b5-immunohistochemistry-human-skin.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [TAL 1B5] - BSA and Azide free (ab176408)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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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [TAL 1B5] - BSA and Azide free (ab176408)](https://www.abcam.com/ps/products/176/ab176408/Images/ab176408-491766-anti-hla-dr-antibody-tal-1b5-immunohistochemistry-human-tonsil.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [TAL 1B5] - BSA and Azide free (ab176408)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 - Anti-HLA-DR antibody [TAL 1B5] - BSA and Azide free (ab176408)](https://www.abcam.com/ps/products/176/ab176408/Images/ab176408-491768-anti-hla-dr-antibody-tal-1b5-flow-cytometry-human-blood.jpg)
Flow Cytometry - Anti-HLA-DR antibody [TAL 1B5] - BSA and Azide free (ab176408)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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (1)
ab176408 has been referenced in 1 publication.
- Schwabenland M et al. Deep spatial profiling of human COVID-19 brains reveals neuroinflammation with distinct microanatomical microglia-T-cell interactions. Immunity 54:1594-1610.e11 (2021). PubMed: 34174183
