Name: Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free
SKU: ab215985
Rating: 5 (1 reviews)

Rabbit Recombinant Monoclonal HLA-DRA antibody. Carrier free. Suitable for ICC/IF, Flow Cyt (Intra), WB, Mass Cytometry, IHC-P and reacts with Human, Transfected cell line - Human samples. Cited in 7 publications.

Images

Immunocytochemistry/ Immunofluorescence - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (AB215985), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	Flow Cyt (Intra)	IP	WB	Mass Cytometry	IHC-P
Human	Tested	Tested	Not recommended	Expected	Tested	Tested
Mouse	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Transfected cell line - Human	Not recommended	Not recommended	Not recommended	Tested	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Transfected cell line - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Transfected cell line - Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Human, Rat, Transfected cell line - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Transfected cell line - Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Transfected cell line - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Transfected cell line - Human	Dilution info -	Notes -

Associated Products

Select an associated product type

15 products for Alternative Product

5 products for Alternative Version

Target data

See full target information HLA-DRA

Function

An alpha chain of antigen-presenting major histocompatibility complex class II (MHCII) molecule. In complex with the beta chain HLA-DRB, displays antigenic peptides on professional antigen presenting cells (APCs) for recognition by alpha-beta T cell receptor (TCR) on HLA-DR-restricted CD4-positive T cells. This guides antigen-specific T-helper effector functions, both antibody-mediated immune response and macrophage activation, to ultimately eliminate the infectious agents and transformed cells (PubMed:15265931, PubMed:15322540, PubMed:17334368, PubMed:22327072, PubMed:24190431, PubMed:27591323, PubMed:29884618, PubMed:31495665, PubMed:8145819, PubMed:9075930). Typically presents extracellular peptide antigens of 10 to 30 amino acids that arise from proteolysis of endocytosed antigens in lysosomes (PubMed:8145819). In the tumor microenvironment, presents antigenic peptides that are primarily generated in tumor-resident APCs likely via phagocytosis of apoptotic tumor cells or macropinocytosis of secreted tumor proteins (PubMed:31495665). Presents peptides derived from intracellular proteins that are trapped in autolysosomes after macroautophagy, a mechanism especially relevant for T cell selection in the thymus and central immune tolerance (PubMed:17182262, PubMed:23783831). The selection of the immunodominant epitopes follows two processing modes: 'bind first, cut/trim later' for pathogen-derived antigenic peptides and 'cut first, bind later' for autoantigens/self-peptides (PubMed:25413013). The anchor residue at position 1 of the peptide N-terminus, usually a large hydrophobic residue, is essential for high affinity interaction with MHCII molecules (PubMed:8145819).

Alternative names

HLA-DRA1, HLA-DRA, MHC class II antigen DRA

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR3692
Purification technique: Affinity purification Protein A
Dissociation constant: 1.67 x 10^-10 M
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab215985 is the carrier-free version of Anti-HLA-DR antibody [EPR3692] ab92511.

Species reactivity
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species.
Please contact us for more information.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

What does low endotoxin mean?
Our low endotoxin, azide-free formats have low endotoxin level (1 EU/mg, determined by the TAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

17 product images

Immunocytochemistry/ Immunofluorescence - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
This ICC/IF data was generated using the same anti-HLA DR antibody clone, EPR3692, in a different buffer formulation (cat# Anti-HLA-DR antibody [EPR3692] ab92511).
Anti-HLA-DR antibody [EPR3692] ab92511 staining HLA-DR in HuT-78 (human Sezary syndrome) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 were used as counterstains for primary antibody Anti-HLA-DR antibody [EPR3692] ab92511 and secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 respectively and DAPI was used as a nuclear counterstain.
Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120)
Negative control 2: Mouse primary antibody (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) and anti-rabbit secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077)
Flow Cytometry (Intracellular) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Overlay histogram showing Raji cells stained with Anti-HLA-DR antibody [EPR3692] ab92511 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Anti-HLA-DR antibody [EPR3692] ab92511, 1/50) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HLA-DR antibody [EPR3692] ab92511).
This image is courtesy of the Single Cell & Imaging Mass Cytometry Analysis Platform, Goodman Cancer Research Centre, McGill University
Mass Cytometry - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Imaging Mass Cytometry™ (IMC™) image of human tonsil tissue stained with Anti-HLA-DR antibody [EPR3692]. ab215985 (carrier-free antibody, purified) was metal-conjugated using a Maxpar® Antibody Labeling Kit from Fluidigm. Immunostaining was performed according to Fluidigm's protocols. Briefly, slides were subject to deparaffinization and heat-induced epitope retrieval, followed by overnight incubation at 4°C with an antibody cocktail containing metal-tagged antibodies in blocking buffer. Slides were subsequently washed with 0.2% Triton-X and 1x PBS, counterstained with Cell-ID™ Intercalator-Ir diluted at 1/400 in 1x PBS for 30 min at room temperature, rinsed for 5 min with distilled H2O, and air-dried prior to IMC™ acquisition. IMC™ acquisition was performed using the Fluidigm Hyperion™ Imaging System.
Imaging Mass Cytometry™, IMC™, Cell-ID™, Hyperion™ and Maxpar® are trademarks of Fluidigm Canada
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Anti-HLA-DR antibody [EPR3692] ab92511 staining HLA-DR in human spleen tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/700. A goat anti-rabbit IgG H&L (HRP) Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.
Negative control 1: PBS in place of primary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HLA-DR antibody [EPR3692] ab92511).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Anti-HLA-DR antibody [EPR3692] ab92511 showing positive staining in Normal liver vessels tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HLA-DR antibody [EPR3692] ab92511).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Anti-HLA-DR antibody [EPR3692] ab92511 showing positive staining in Normal skin vessels tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HLA-DR antibody [EPR3692] ab92511).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Anti-HLA-DR antibody [EPR3692] ab92511 showing positive staining in Endometrial carcinoma vessels tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HLA-DR antibody [EPR3692] ab92511).
OI-RD Scanning - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labelling HLA-DR with Anti-HLA-DR antibody [EPR3692] ab92511 at 1/10000 dilution (0.07 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) secondary antibody. Positive staining on human tonsil tissue. The section was incubated with Anti-HLA-DR antibody [EPR3692] ab92511 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control.
Heat mediated antigen retrieval with Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 30 minutes.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HLA-DR antibody [EPR3692] ab92511).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Immunohistochemical analysis of paraffin-embedded human skin tissue labelling HLA-DR with Anti-HLA-DR antibody [EPR3692] ab92511 at 1/10000 dilution (0.07 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) secondary antibody. Positive staining on human skin tissue. The section was incubated with Anti-HLA-DR antibody [EPR3692] ab92511 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control.
Heat mediated antigen retrieval with Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 30 minutes.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HLA-DR antibody [EPR3692] ab92511).
Western blot - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Blocking buffer and concentration : 5% NFDM/TBST
Diluting buffer and concentration : 5% NFDM/TBST
This antibody does not cross-react with human HLA-DOA.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HLA-DR antibody [EPR3692] ab92511).
Lanes 1 - 2: Western blot - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985) at 1/1000 dilution
Lanes 1 - 2: Western blot - Anti-HLA-DR antibody [EPR3692] - BSA and Azide free (Anti-HLA-DR antibody [EPR3692] - BSA and Azide free ab209968) at 1/1000 dilution
Lanes 1 - 2: Western blot - Anti-HLA-DR antibody [EPR3692] (Anti-HLA-DR antibody [EPR3692] ab92511) at 1/1000 dilution
Lane 1: HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with a human HLA-DR expression vector containing a his-tag, whole cell lysate at 20 µg
Lane 2: HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with a human HLA-DOA expression vector containing a his-tag, whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 29 kDa
Observed band size: 37 kDa
Exposure time: 3min
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Tissue Microarrays stained for "Anti-HLA-DR antibody [EPR3692]" using "Anti-HLA-DR antibody [EPR3692] ab92511"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with Anti-HLA-DR antibody [EPR3692] ab92511 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Anti-HLA-DR antibody [EPR3692] ab92511, at a 1/100 dilution, staining HLA-DRA in paraffin embedded Human tonsil (A) and Human kidney (B) tissue by Immunohistochemistry. Detection: by DAB staining
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HLA-DR antibody [EPR3692] ab92511).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Anti-HLA-DR antibody [EPR3692] ab92511 showing positive staining in Endometrial carcinoma vessels tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HLA-DR antibody [EPR3692] ab92511).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Anti-HLA-DR antibody [EPR3692] ab92511 showing positive staining in Normal skin vessels tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HLA-DR antibody [EPR3692] ab92511).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Anti-HLA-DR antibody [EPR3692] ab92511 showing positive staining in Normal liver vessels tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HLA-DR antibody [EPR3692] ab92511).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)
Anti-HLA-DR antibody [EPR3692] ab92511 staining HLA-DR in human spleen tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/700. A goat anti-rabbit IgG H&L (HRP) Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1: PBS in place of primary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HLA-DR antibody [EPR3692] ab92511).