PSGL-1 Recombinant Rabbit Monoclonal Antibody [PSH20-00]
cat.: HA724120
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Mouse, Rat
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: PSH20-00
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer: 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 42 kDa
Isotype: IgG
Positive control: Mouse thymus tissue lysate, Mouse spleen tissue lysate, Mouse kidney tissue lysate, Rat spleen tissue lysate, Rat kidney tissue lysate, RAW264.7 cell lysate, A20 cell lysate, NIH/3T3 cell lysate, mouse splenocyte, mouse colon tissue, mouse spleen tissue, rat colon tissue, rat spleen tissue.
Subcellular location: Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
1:5,000
1:500
1:1,000
Uniprot #: SwissProt: Q62170 Mouse
Entrez Gene: 363930 Rat
Alternative names: CD 162 CD162 CD162 antigen CLA Cutaneous lymphocyte associated associated antigen P selectin glycoprotein ligand 1 P selectin glycoprotein ligand 1 precursor P-selectin glycoprotein ligand 1 PSGL 1 PSGL-1 PSGL1 Selectin P ligand SELPLG
Images
HA724120_1.jpg Fig1: Western blot analysis of PSGL-1 on different lysates with Rabbit anti-PSGL-1 antibody (HA724120) at 1/5,000 dilution.

Lane 1: Mouse thymus tissue lysate (40 µg/Lane)
Lane 2: Mouse spleen tissue lysate (40 µg/Lane)
Lane 3: Mouse kidney tissue lysate (low expression) (40 µg/Lane)
Lane 4: Rat spleen tissue lysate (40 µg/Lane)
Lane 5: Rat kidney tissue lysate (low expression) (40 µg/Lane)

Predicted band size: 42 kDa
Observed band size: 100-200 kDa

Exposure time: 20 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 (HA724120) 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.
HA724120_2.jpg Fig2: Western blot analysis of PSGL-1 on different lysates with Rabbit anti-PSGL-1 antibody (HA724120) at 1/5,000 dilution.

Lane 1: RAW264.7 cell lysate (20 µg/Lane)
Lane 2: A20 cell lysate (20 µg/Lane)
Lane 3: NIH/3T3 cell lysate (low expression) (20 µg/Lane)

Predicted band size: 42 kDa
Observed band size: 100 kDa

Exposure time: 3 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 (HA724120) 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.
HA724120_3.jpg Fig3: Immunocytochemistry analysis of mouse splenocyte labeling PSGL-1 with Rabbit anti-PSGL-1 antibody (HA724120) at 1/500 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-PSGL-1 antibody (HA724120) at 1/500 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.
HA724120_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-PSGL-1 antibody (HA724120) 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 (HA724120) 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.
HA724120_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-PSGL-1 antibody (HA724120) 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 (HA724120) 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.
HA724120_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue (low expression) with Rabbit anti-PSGL-1 antibody (HA724120) 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 (HA724120) 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.
HA724120_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-PSGL-1 antibody (HA724120) 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 (HA724120) 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.
HA724120_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-PSGL-1 antibody (HA724120) 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 (HA724120) 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.
HA724120_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue (low expression) with Rabbit anti-PSGL-1 antibody (HA724120) 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 (HA724120) 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.
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