Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (AB215985)

This data was developed using ab92511 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling HLA-DR with ab92511 at a concentration of 0.5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32 mins. ab209968 Anti-HLA-DR antibody [EPR3692] was incubated for 16 mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• Mass Cytometry

Collaborator

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

This image is courtesy of the Single Cell & Imaging Mass Cytometry Analysis Platform, Goodman Cancer Research Centre, McGill University

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (AB215985)

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) 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 (ab92511).

• ICC/IF

Supplier Data

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# ab92511).

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) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab7291 and ab150120 were used as counterstains for primary antibody ab92511 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

Negative control 1 : Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
Negative control 2 : Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (AB215985)

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 (ab92511).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (AB215985)

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 (ab92511).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (AB215985)

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 (ab92511).

• IHC-P

Lab

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 "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 ab92511 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (AB215985)

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) 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 (ab92511).

• mIHC

Collaborator

Multiplex immunohistochemistry - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (AB215985)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).

10-color fluorescence multiplex immunohistochemical analysis of human lung cancer tissue (formalin-fixed paraffin-embedded section).

Merged staining of anti-FOXP3 (ab215206; Cyan; TG540N), anti-PD1 (ab52587; Red; TG700N), anti-CD163 (ab182422; Brown; TG650N), anti-HLA-DR (ab92511; Yellow; TG570N), anti-CD4 (ab133616; Violet; TG620N), anti-CD8 alpha (ab101500; Purple; TG540S), anti-CD20 (ab9475; Grey; TG660S), anti-CD68 (ab192847; Green; TG520N), anti-Cytokeratin 19 (ab52625; Light blue; TG440N). TG470SN (dark blue) was used as a nuclear counter stain. The inset image shows the separate CD68 signal.

The section was incubated in nine rounds of staining; in the order of ab215206 (1/100 dilution), ab52587 (1/200 dilution), ab182422 (1/300 dilution), ab92511 (1/200 dilution), ab133616 (1/600 dilution), ab101500 (1/300 dilution), ab9475 (1/100 dilution), ab192847 (1/300 dilution), ab52625 (1/400 dilution); each using a separate fluorescent tyramide signal amplification system.

Sodium citrate antigen retrieval (pH6.0) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

Image acquisition was performed with TissueFAXS Spectra (TissueGnostics).

This image is courtesy of TissueGnostics Asia Pacific Limited

• IHC-P

Lab

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 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 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 (ab92511).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (AB215985)

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 (ab92511).

• IHC-P

Lab

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 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 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 (ab92511).

• WB

Lab

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 (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 (<a href='/en-us/products/primary-antibodies/hla-dr-antibody-epr3692-bsa-and-azide-free-ab209968'>ab209968</a>) at 1/1000 dilution

Lanes 1 - 2:

Western blot - Anti-HLA-DR antibody [EPR3692] (<a href='/en-us/products/primary-antibodies/hla-dr-antibody-epr3692-ab92511'>ab92511</a>) 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 29 kDa

Observed band size: 37 kDa

false

Exposure time: 3min

• OI-RD Scanning

Unknown

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.