Thrombomodulin Recombinant Rabbit Monoclonal Antibody [PSH23-10]
cat.: HA724350
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: PSH23-10
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: 60 kDa
Isotype: IgG
Immunogen: Recombinant protein within human Thrombomodulin aa 201-515.
Subcellular location: Membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:5,000-1:20,000
1:500
1:200-1:1,000
1:10,000
1-2μg/sample
Uniprot #: SwissProt: P07204 Human
Alternative names: AHUS 6 AHUS6 BDCA 3 BDCA3 CD 141 CD141 CD141 antigen Fetomodulin Thbd THPH12 THRM Thrombomodulin TM TRBM_HUMAN
Images
HA724350_1.jpg Fig1: Western blot analysis of Thrombomodulin on different lysates with Rabbit anti-Thrombomodulin antibody (HA724350) at 1/20,000 dilution.

Lane 1: A431 (Human epidermoid carcinoma skin squamous cells) cell lysate
Lane 2: 293T (Human embryonic kidney cells) cell lysate
Lane 3: THP-1 (Human acute monoblastic leukemia cells) cell lysate
Lane 4: Jurkat (Human T-lymphoblastic cells) cell lysate

Lysates/proteins at 10 µg/Lane.
Exposure time: 2 minutes; ECL: K1801

293T is negative control (PMID: 40414460).
Negative expression of Thrombomodulin protein in Jurkat cell is consistent with the predicted expression pattern.

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724350, 1/20,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 60 kDa
Observed band size: 75-100 kDa
HA724350_2.jpg Fig2: Western blot analysis of Thrombomodulin on different lysates with Rabbit anti-Thrombomodulin antibody (HA724350) at 1/5,000 dilution.

Lane 1: A431 (Human epidermoid carcinoma skin squamous cells) cell lysate
Lane 2: A431 treated with PNGase F cell lysate

Lysates/proteins at 10 µg/Lane.
Exposure time: 1 minute; ECL: K1801

Thrombomodulin is a glycoprotein and can be deglycosylated by PNGase F. The molecular mass observed is consistent with the literature (PMID: 2827310)

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724350, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 60 kDa
Observed band size: 75-100 kDa
HA724350_3.jpg Fig3: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: A431 (Human epidermoid carcinoma skin squamous cells)/293T (Human embryonic kidney cells)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.

Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA724350, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.

Counterstain: Beta tubulin (HA601187, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).
Negative control: 293T (PMID: 40414460).
HA724350_4.jpg Fig4: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Placenta
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724350, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724350_5.jpg Fig5: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: squamous cell lung carcinoma
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724350, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724350_6.jpg Fig6: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Lymph node
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724350, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724350_7.jpg Fig7: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Esophagus
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724350, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724350_8.jpg Fig8: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Ovarian carcinoma
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724350, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724350_9.jpg Fig9: Application: Flow Cytometry

Species: Human
Sample: A431 (Human epidermoid carcinoma skin squamous cells)/293T (Human embryonic kidney cells)

Blocking: 1% BSA + 10% normal goat serum + human FcR (Miltenyi Biotec), 15 minutes at room temperature. (human)
Antibody dilution buffer: 1x PBS.
Primary antibody: HA724350 (1/10,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
Negative control: 293T (PMID: 40414460).
HA724350_10.jpg Fig10: Application: Flow Cytometry

Species: Human
Sample: A431 (Human epidermoid carcinoma skin squamous cells)/Jurkat (Human T-lymphoblastic cells)

Blocking: 1% BSA + 10% normal goat serum + human FcR (Miltenyi Biotec), 15 minutes at room temperature. (human)
Antibody dilution buffer: 1x PBS.
Primary antibody: HA724350 (1/10,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
Negative expression of Thrombomodulin protein in Jurkat cell is consistent with the predicted expression pattern.
HA724350_11.jpg Fig11: Immunoprecipitation (IP)

Thrombomodulin was immunoprecipitated in 0.2 mg THP-1 (Human acute monoblastic leukemia cells) cell lysate with HA724350 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724350 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: THP-1 cell lysate (input)
Lane 2: HA724350 IP in THP-1 cell lysate
Lane 3: Rabbit IgG instead of HA724350 in THP-1 cell lysate

Exposure time: 8 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA724350, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 60 kDa
Observed band size: 75-100 kDa
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.