PRODUCT CODE: ET1610-66

HLA-DR Recombinant Rabbit Monoclonal Antibody [SC06-78] (ET1610-66)

  • Recombinant

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of HLA-DR on Daudi cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-66, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of HLA-DR on Daudi cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-66, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • ICC staining of HLA-DR in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of HLA-DR in B16F1 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-HLA-DR antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-66, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-HLA-DR antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-66, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-HLA-DR antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-66, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of HLA-DR was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1610-66, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of HLA-DR on Daudi cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-66, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

HLA-DR Recombinant Rabbit Monoclonal Antibody [SC06-78] (ET1610-66)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

Daudi cell lysates, Hela, B16F1, human tonsil tissue, human liver tissue, human spleen tissue, Jurkat.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SC06-78

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

29 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:5,000

  • ICC

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

HLA-DR

SYNONYMS

DR alpha chain antibody; DR alpha chain precursor antibody; DRA_HUMAN antibody; DRB1 antibody; DRB4 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-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

SEQUENCE SIMILARITIES

Belongs to the MHC class II family.

POST-TRANSLATIONAL MODIFICATION

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.

SUBCELLULAR LOCATION

Cell membrane, Endoplasmic reticulum membrane, Golgi apparatus, Endosome membrane, Lysosome membrane.

FUNCTION

Major histocompatibility complex (MHC) class II molecules destined for presentation to CD4+ helper T cells is determined by two key events. These events include the dissociation of class II-associated invariant chain peptides (CLIP) from an antigen binding groove in MHC II α/β dimers through the activity of MHC molecules HLA-DM and -DO, and subsequent peptide antigen binding. Accumulating in endosomal/lysosomal compartments and on the surface of B cells, HLA-DM, -DO molecules regulate the dissociation of CLIP and the subsequent binding of exogenous peptides to HLA class II molecules (HLA-DR, -DQ and -DP) by sustaining a conformation that favors peptide exchange. RFLP analysis of HLA-DM genes from rheumatoid arthritis (RA) patients suggests that certain polymorphisms are genetic factors for RA susceptibility. HLA-B belongs to the HLA class I heavy chain paralogs. Class I molecules play a central role in the immune system by presenting peptides derived from the endoplasmic reticulum lumen. HLA-B and -C can form heterodimers consisting of a membrane anchored heavy chain and a light chain (β-2-Microglobulin). Polymorphisms yield hundreds of HLA-B and -C alleles.