HLA-ABC (MHC I) Recombinant Rabbit Monoclonal Antibody [PSH12-02]
cat.: HA723415
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: PSH12-02
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 40 kDa
Isotype: IgG
Immunogen: Recombinant protein within human HLA-B aa 110-309.
Positive control: Raji cell lysate, THP-1 cell lysate, Jurkat cell lysate, HL-60 cell lysate, HepG2 cell lysate, Ramos cell lysate, Mouse spleen tissue lysate, Mouse lung tissue lysate, Rat spleen tissue lysate, Rat lung tissue lysate, Jurkat, Raji, human spleen tissue, human stomach tissue, mouse spleen tissue, mouse stomach tissue, rat spleen tissue, rat stomach tissue.
Subcellular location: Cell membrane, Endoplasmic reticulum membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
1:5,000
1:100
1:2,000
Uniprot #: SwissProt: P04439 Human | P01889 Human | P10321 Human
Alternative names: Antigen Presenting Molecule Cw*1701 D6S204 FLJ27082 HLA A HLA B HLA C HLA class 1 A HLA class 1 B HLA class 1 C HLA JY3 HLAA HLAB HLAC HLC C Human Leucocyte Antigen C Major Histocompatibility Complex Class I A Major Histocompatibility Complex Class I B Major Histocompatibility Complex Class I C Major histocompatibility complex, class I, A + B + C MHC class I antigen A*1 MHC Class I Antigen MHC class I antigen B*7 MHC class I antigen Cw*1 MHC class I HLA A MHC class I HLA B MHC class I HLA C MHC HLA ABC PSORS1
Images
HA723415_1.jpg Fig1: Western blot analysis of HLA-ABC (MHC I) on different lysates with Rabbit anti-HLA-ABC (MHC I) antibody (HA723415) at 1/5,000 dilution.

Lane 1: Raji cell lysate (20 µg/Lane)
Lane 2: THP-1 cell lysate (20 µg/Lane)
Lane 3: Jurkat cell lysate (20 µg/Lane)
Lane 4: HL-60 cell lysate (20 µg/Lane)
Lane 5: HepG2 cell lysate (20 µg/Lane)
Lane 6: Ramos cell lysate (20 µg/Lane)
Lane 7: Mouse spleen tissue lysate (40 µg/Lane)
Lane 8: Mouse lung tissue lysate (40 µg/Lane)
Lane 9: Rat spleen tissue lysate (40 µg/Lane)
Lane 10: Rat lung tissue lysate (40 µg/Lane)

Predicted band size: 40 kDa
Observed band size: 45 kDa

Exposure time: 10 seconds; 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 (HA723415) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA723415_2.jpg Fig2: Immunocytochemistry analysis of Jurkat cells labeling HLA-ABC (MHC I) with Rabbit anti-HLA-ABC (MHC I) antibody (HA723415) at 1/100 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-HLA-ABC (MHC I) antibody (HA723415) at 1/100 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.
HA723415_3.jpg Fig3: Immunocytochemistry analysis of Raji cells labeling HLA-ABC (MHC I) with Rabbit anti-HLA-ABC (MHC I) antibody (HA723415) at 1/100 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-HLA-ABC (MHC I) antibody (HA723415) at 1/100 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.
HA723415_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-HLA-ABC (MHC I) antibody (HA723415) at 1/2,000 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 (HA723415) at 1/2,000 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.
HA723415_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-HLA-ABC (MHC I) antibody (HA723415) at 1/2,000 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 (HA723415) at 1/2,000 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.
HA723415_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-HLA-ABC (MHC I) antibody (HA723415) at 1/2,000 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 (HA723415) at 1/2,000 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.
HA723415_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-HLA-ABC (MHC I) antibody (HA723415) at 1/2,000 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 (HA723415) at 1/2,000 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.
HA723415_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-HLA-ABC (MHC I) antibody (HA723415) at 1/2,000 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 (HA723415) at 1/2,000 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.
HA723415_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat stomach tissue with Rabbit anti-HLA-ABC (MHC I) antibody (HA723415) at 1/2,000 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 (HA723415) at 1/2,000 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.
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