Name: Anti-HLA-DR antibody [EPR3692]
Brand: RabMAb
SKU: ab92511
Rating: 4 (7 reviews)

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Multiplex immunohistochemistry - Anti-HLA-DR antibody [EPR3692] (ab92511)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR3692] to HLA-DR
Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IF, mIHC
Reacts with: Rat, Human

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 647 Carrier Free

Product name

Anti-HLA-DR antibody [EPR3692]
See all HLA-DR primary antibodies
Description

Rabbit monoclonal [EPR3692] to HLA-DR
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IF, mIHCmore details
Unsuitable for: IP
Species reactivity

Reacts with: Rat, Human
Does not react with: Mouse
Immunogen

Synthetic peptide within Human HLA-DR aa 150-250. The exact sequence is proprietary.
Database link: P01903
Positive control
- WB: Raji, Human spleen and Hu T-78 cell lysates, Ramos whole cell lysate, Human tonsil tissue and HEK-293T cells transfected with a human HLA-DR expression vector containing a his-tag whole cell lysate. ICC/IF: HuT-78 cells. Flow Cyt (Intra): Raji cells. IHC-P: Human tonsil tissue, human skin tissue, human spleen tissue, liver vessels tissue, skin vessels tissue, endometrial carcinoma vessels tissue, human kidney tissue, rat spleen tissue and rat colon tissue. mIHC: Hu lung cancer tissue
General notes
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Dissociation constant (K_D)

K_D = 1.67 x 10 ^-10 M
Learn more about K_D
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR3692
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human HLA-DR protein (ab177661)
Related Products
- Anti-HLA-DP antibody [B7/21] (ab20897)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab92511 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		1/1000 - 1/10000. Predicted molecular weight: 29 kDa.
IHC-P	(4)	1/100 - 1/10000.
Flow Cyt (Intra)		1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF		1/250 - 1/500.
mIHC		1/200.

Notes
WB 1/1000 - 1/10000. Predicted molecular weight: 29 kDa.
IHC-P 1/100 - 1/10000.
Flow Cyt (Intra) 1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF 1/250 - 1/500.
mIHC 1/200.

Application notes

Is unsuitable for IP.

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
modifications

Ubiquitinated 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.
Target information above from: UniProt accession P01903 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3122 Human
- Omim: 142860 Human
- SwissProt: P01903 Human
- Unigene: 520048 Human
Alternative names
- DASS-397D15.1 antibody
- DR alpha chain antibody
- DR alpha chain precursor antibody
- DRA_HUMAN antibody
- DRB1 antibody
- DRB4 antibody
- FLJ51114 antibody
- Histocompatibility antigen HLA DR alpha antibody
- Histocompatibility antigen HLA-DR alpha antibody
- HLA class II histocompatibility antigen antibody
- HLA class II histocompatibility antigen DR alpha chain antibody
- HLA DR1B antibody
- HLA DR3B antibody
- HLA DRA antibody
- HLA DRA1 antibody
- HLA DRB1 antibody
- HLA DRB3 antibody
- HLA DRB4 antibody
- HLA DRB5 antibody
- HLA-DR histocompatibility type antibody
- HLA-DRA antibody
- HLADR4B antibody
- HLADRA1 antibody
- HLADRB antibody
- Major histocompatibility complex class II DR alpha antibody
- Major histocompatibility complex class II DR beta 1 antibody
- Major histocompatibility complex class II DR beta 3 antibody
- Major histocompatibility complex class II DR beta 4 antibody
- Major histocompatibility complex class II DR beta 5 antibody
- MGC117330 antibody
- MHC cell surface glycoprotein antibody
- MHC class II antigen DRA antibody
- MHC II antibody
- MLRW antibody
- OTTHUMP00000029406 antibody
- OTTHUMP00000029407 antibody
see all

Multiplex immunohistochemistry - Anti-HLA-DR antibody [EPR3692] (ab92511)This image is courtesy of TissueGnostics Asia Pacific Limited

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 HLA-DR 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).
Multiplex immunohistochemistry - Anti-HLA-DR antibody [EPR3692] (ab92511)This image is courtesy of TissueGnostics Asia Pacific Limited

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; Violet; TG700N), anti-CD163 (ab182422; Red; TG650N), anti-HLA-DR (ab92511; Yellow; TG570N), anti-CD4 (ab133616; Orange; 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 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).
Western blot - Anti-HLA-DR antibody [EPR3692] (ab92511)

All lanes : 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
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

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 29 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking buffer and concentration : 5% NFDM/TBST
Diluting buffer and concentration : 5% NFDM/TBST

This antibody does not cross-react with human HLA-DOA.
Immunocytochemistry/ Immunofluorescence - Anti-HLA-DR antibody [EPR3692] (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)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] (ab92511)

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.
Flow Cytometry (Intracellular) - Anti-HLA-DR antibody [EPR3692] (ab92511)

Overlay histogram showing Raji cells stained with 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 (ab92511, 1/50) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (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.
Western blot - Anti-HLA-DR antibody [EPR3692] (ab92511)

All lanes : Anti-HLA-DR antibody [EPR3692] (ab92511) at 1/10000 dilution

Lane 1 : Raji (human Burkitt's lymphoma) whole cell lysate
Lane 2 : Ramos (human Burkitt's lymphoma) whole cell lysate
Lane 3 : Human tonsil tissue

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 29 kDa

Blocking buffer: 5% NFDM/TBST

Diluting buffer: 5% NFDM/TBST
Western blot - Anti-HLA-DR antibody [EPR3692] (ab92511)

All lanes : Anti-HLA-DR antibody [EPR3692] (ab92511) at 1/5000 dilution

Lane 1 : Raji cell lysate
Lane 2 : Human spleen lysate
Lane 3 : Hu T-78 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 29 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] (ab92511)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] (ab92511)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] (ab92511)

ab92511 showing positive staining in Normal liver vessels tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] (ab92511)

ab92511 showing positive staining in Normal skin vessels tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] (ab92511)

ab92511 showing positive staining in Endometrial carcinoma vessels tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] (ab92511)

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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] (ab92511)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling HLA DR with ab92511 at a dilution of 1/1000. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody, at a dilution of 1/500. Counter stained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] (ab92511)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling HLA DR with ab92511 at a dilution of 1/1000. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody, at a dilution of 1/500. Counter stained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [EPR3692] (ab92511)

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.
OI-RD Scanning - Anti-HLA-DR antibody [EPR3692] (ab92511)

Equilibrium disassociation constant (K_D)
Learn more about K_D

Click here to learn more about K_D
Anti-HLA-DR antibody [EPR3692] (ab92511)

Flow cytometry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab92511? Please let us know so that we can cite the reference in this datasheet.

ab92511 has been referenced in 23 publications.

Wang Z et al. Polarization of intestinal tumour-associated macrophages regulates the development of schistosomal colorectal cancer. J Cancer 12:1033-1041 (2021). PubMed: 33442402
Kong X et al. Potential Role of Macrophage Phenotypes and CCL2 in the Pathogenesis of Takayasu Arteritis. Front Immunol 12:646516 (2021). PubMed: 34079541
Duan R et al. Tumor Microenvironment Status Predicts the Efficacy of Postoperative Chemotherapy or Radiochemotherapy in Resected Gastric Cancer. Front Immunol 11:609337 (2020). PubMed: 33569057
Pan C et al. Pretreatment with human urine-derived stem cells protects neurological function in rats following cardiopulmonary resuscitation after cardiac arrest. Exp Ther Med 20:112 (2020). PubMed: 32989390
Cowman SJ et al. Macrophage HIF-1a Is an Independent Prognostic Indicator in Kidney Cancer. Clin Cancer Res 26:4970-4982 (2020). PubMed: 32586940

View all Publications for this product

Customer reviews and Q&As

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1-8 of 8 Abreviews or Q&A

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Cynomolgus monkey Tissue sections (Mesenteric Lymph Node)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: pH 9

Permeabilization

Specification

Mesenteric Lymph Node

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 22°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Blaise LAFOUX

Verified customer

Submitted Oct 11 2022

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Colorectal cancer)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrate pH 6

Permeabilization

Specification

Colorectal cancer

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 02 2021

Application

IHC - Wholemount

Sample

Human Tissue (Kidney)

Specification

Kidney

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Daniyal Jafree

Verified customer

Submitted May 28 2021

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (NSCLC)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citritc acid pH6

Permeabilization

Specification

NSCLC

Blocking step

Zytomed Systems Antibody Diluent as blocking agent for 10 minute(s) · Concentration: 100% · Temperature: 21°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 12 2020

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Meninges)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH8

Permeabilization

Yes - 0.1% Tween in PBS

Specification

Meninges

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 20°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 16 2019

Application

Flow Cytometry

Sample

Rabbit Cell (adipose)

Permeabilization

Specification

adipose

Fixation

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 20 2018

Application

Immunohistochemistry (Frozen sections)

Sample

Human Tissue sections (Liver)

Specification

Liver

Fixative

Formaldehyde

Permeabilization

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 27°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 08 2011

Question

Inquiry: Dear Sir or Madam: We recently purchased the Anti-HLA-DR antibody [EPR3692] ab92511 from Abcam. We tested the antibody in IHC using normal human skin as positive control. However, the antibody only detected basal keratinocytes but not immune cells which typically show strong HLA-DR staining. Importantly, our IHC procedure works fine for other antibodies, e.g. Langerin with detects Langerhans cells highly specific, but we could not get any specific signal for the HLA-DR antibody. Only basal keratinocytes were detected (see attached picture). Species: human skin (frozen 5 µm sections) Sample fixation: ice-cold acetone Blocking conditions: 5% goat serum (in PBS) for 1h at room temperature Detection method: MaxPoly-Two Plus Polymer HRP (Dianova) Primary antibody conditions: dilution 1:250 (in PBS/ 0.0025% BSA/ 0.025% Tween), 1h incubation at room temperature Secondary antibody conditions: MaxPoly-Two Plus Polymer HRP Universal Detection Kit (according to instructions) Wash steps: with PBS/ 0.0025% BSA/ 0.025% Tween
If possible we would like to test another HLA-DR antibody for our purposes (e.g Anti-HLA DR antibody [TAL 1B5] (ab20181)) although we would prefer a rabbit antibody. Thank you very much in advance.

Abcam community

Verified customer

Asked on Oct 26 2012

Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I would like to reassure you that ab92511 is tested and covered by our 6 month guarantee for use in IHC-P and human samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would like to offer some suggestions to help optimise the results from ab92511

We have not received any other complaints for this lot.

I suspect that the issue may be the fixation method, as we have only confirmed the antibody to work with formalin fixed/paraffin embedded tissue. I would suggest that to repeat the experiment using our recommended protocol (http://www.epitomics.com/products/support/ihc), including increasing the primary dilution up to 1:100 and increasing the incubation time to overnight.

If you have not used the same secondary system recently with another rabbit primary I would also suggest verifying that it is working properly.

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Abcam Scientific Support

Answered on Oct 26 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Dissociation constant (KD)

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (23)

Customer reviews and Q&As

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HLA-DR antibody [EPR3692]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HLA-DR antibody [EPR3692]

IHC - Wholemount abreview for Anti-HLA-DR antibody [EPR3692]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HLA-DR antibody [EPR3692]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HLA-DR antibody [EPR3692]

Flow Cytometry abreview for Anti-HLA-DR antibody [EPR3692]

Immunohistochemistry (Frozen sections) abreview for Anti-HLA-DR antibody [EPR3692]

Dissociation constant (K_D)

Post-translational
modifications