ET1610-66

HLA-DR Recombinant Rabbit Monoclonal Antibody [SC06-78]

cat.: ET1610-66

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	SC06-78

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 29 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human HLA-DRA aa 159-195 (Extracellular).
Positive control:	Daudi cell lysates, Raji cell lysates, Hela, B16F1, human tonsil tissue, human liver tissue, human spleen tissue, Jurkat.
Subcellular location:	Cell membrane, Endoplasmic reticulum membrane, Golgi apparatus, Endosome membrane, Lysosome membrane.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:1,000 1:50-1:200 1:50-1:200 1:50-1:100
Uniprot #:	SwissProt: P01903 Human
Alternative names:	DR alpha chain DR alpha chain precursor DRA_HUMAN DRB1 DRB4 Histocompatibility antigen HLA DR alpha HLA class II histocompatibility antigen HLA class II histocompatibility antigen DR alpha chain HLA DR1B HLA DR3B HLA DRA HLA DRA1 HLA DRB1 HLA DRB3 HLA DRB4 HLA DRB5 HLA-DRA HLADR4B HLADRA1 HLADRB Major histocompatibility complex class II DR alpha Major histocompatibility complex class II DR beta 1 Major histocompatibility complex class II DR beta 3 Major histocompatibility complex class II DR beta 4 Major histocompatibility complex class II DR beta 5 MGC117330 MHC cell surface glycoprotein MHC class II antigen DRA MHC II MLRW

Images

Fig1: Western blot analysis of HLA-DR on different lysates with Rabbit anti-HLA-DR antibody (ET1610-66) at 1/1,000 dilution.

Lane 1: Daudi cell lysate
Lane 2: Raji cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 29 kDa
Observed band size: 35 kDa

Exposure time: 1 minute 40 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-66) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of Hela cells labeling HLA-DR with Rabbit anti-HLA-DR antibody (ET1610-66) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-HLA-DR antibody (ET1610-66) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

	Fig3: Immunocytochemistry analysis of B16F1 cells labeling HLA-DR with Rabbit anti-HLA-DR antibody (ET1610-66) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-HLA-DR antibody (ET1610-66) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
	Fig4: 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 9.0) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-HLA-DR antibody (ET1610-66) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1610-66) at 1/1,000 dilution for 1 hour 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.
	Fig6: 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 9.0) 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.

Fig7: 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).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.