MHC Class II Recombinant Rabbit Monoclonal Antibody [JA10-94]
cat.: ET1704-13
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IP, IF-Cell, IF-Tissue, IHC-P, mIHC
Clonality: Monoclonal
Clone number: JA10-94
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
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-DPB1 aa 11-60 / 258.
Positive control: Daudi cell lysates, human tonsil tissue, human endometrium tissue, Daudi.
Subcellular location: Cell membrane. Endoplasmic reticulum membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  IP
  mIHC
1:5,000
1:50
1:50-1:100
1:1,000
Use at an assay dependent concentration.
1:2,000
Uniprot #: SwissProt: P04440 Human
Alternative names: D6S221E DMA DMB DP beta 1 DP beta 1 chain DP(W4) beta chain DPB 1 DPB1 DPB1_HUMAN DRB H2Ea HLA class II histocompatibility antigen HLA class II histocompatibility antigen DM beta chain HLA class II histocompatibility antigen, DP beta 1 chain HLA class II histocompatibility antigen, DP(W4) beta chain HLA DMB HLA DP1A HLA DPB1 HLA SB alpha chain HLA-A HLA-A histocompatibility type HLA-DP HLA-DP histocompatibility type, beta-1 subunit HLA-DP1B HLA-DPB HLA-DPB1 HLADM HLADP1B HLASB HLASB histocompatibility type Human MHC class II HLA SB alpha LA class II histocompatibility antigen DP alpha 1 chain Major histocompatibility complex class II Major histocompatibility complex class II DP alpha 1 Major histocompatibility complex class II DP beta 1 Major histocompatibility complex, class I, A MHC class II antigen DMB MHC class II antigen DPB1 MHC class II DP3 alpha MHC class II DPA1 MHC class II HLA-DP-beta-1 MHC DPB......
Images
ET1704-13_1.jpg Fig1: Western blot analysis of MHC Class II on Daudi cell lysates with Rabbit anti-MHC Class II antibody (ET1704-13) at 1/5,000 dilution.

Lysates/proteins at 120 µg/Lane.

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

Exposure time: 2 minutes; ECL: K1801;

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 (ET1704-13) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1704-13_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-MHC Class II antibody (ET1704-13) 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 (ET1704-13) 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.
ET1704-13_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human endometrium tissue with Rabbit anti-MHC Class II antibody (ET1704-13) 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 (ET1704-13) 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.
ET1704-13_4.jpg Fig4: Immunocytochemistry analysis of Daudi cells labeling MHC Class II with Rabbit anti-MHC Class II antibody (ET1704-13) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MHC Class II antibody (ET1704-13) at 1/50 dilution in 1% BSA in PBST 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.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1704-13_5.jpg Fig5: Fluorescence multiplex immunohistochemical analysis of human tonsil (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-HLA-DBP1 (ET1704-13, Green) and anti-Tryptase (ET1610-64, Red) on tonsil. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in two rounds of staining: in the order of ET1704-13 (1/2,000 dilution) and ET1610-64 (1/5,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Zeiss Observer 7 Inverted Fluorescence Microscope.
ET1704-13_6.jpg Fig6: Fluorescence multiplex immunohistochemical analysis of Human tonsil (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-BCL6 (HA601083, Red), anti-HLA-DPB1 (ET1704-13, Green), anti-Tryptase (ET1610-64, White), anti-CD20 (HA721138, Magenta) and anti-CD45 (ET7111-03, Yellow) on tonsil. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in five rounds of staining: in the order of HA601083 (1/200 dilution), ET1704-13 (1/2,000 dilution), ET1610-64 (1/5,000 dilution), HA721138 (1/2,000 dilution) and ET7111-03 (1/500 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.