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Thrombomodulin Recombinant Rabbit Monoclonal Antibody [PSH23-10] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751888

Thrombomodulin Recombinant Rabbit Monoclonal Antibody [PSH23-10] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

  • IP

Reactivity

  • Human

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

Thrombomodulin Recombinant Rabbit Monoclonal Antibody [PSH23-10] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human Thrombomodulin aa 201-515.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IHC-P, FC, IP

Molecular Weight

Predicted band size: 60 kDa

Conjugation

unconjugated

Clone Number

PSH23-10

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000-1:20,000

  • IF-Cell

  • 1:500

  • IHC-P

  • 1:200-1:1,000

  • FC

  • 1:10,000

  • IP

  • 1-2μg/sample

Target

Function

Thrombomodulin functions as a cofactor in the thrombin-induced activation of protein C in the anticoagulant pathway by forming a 1:1 stoichiometric complex with thrombin. This raises the speed of protein C activation thousandfold. Thrombomodulin-bound thrombin has procoagulant effect at the same time by inhibiting fibrinolysis by cleaving thrombin-activatable fibrinolysis inhibitor (TAFI, aka carboxypeptidase B2) into its active form. Thrombomodulin is a glycoprotein on the surface of endothelial cells that, in addition to binding thrombin, regulates C3b inactivation by factor I. Mutations in the thrombomodulin gene (THBD) have also been reported to be associated with atypical hemolytic-uremic syndrome (aHUS).

Background References

1. Giri H et al. Thrombomodulin: a multifunctional receptor modulating the endothelial quiescence. J Thromb Haemost. 2024 Apr

2. Boron M et al. Circulating Thrombomodulin: Release Mechanisms, Measurements, and Levels in Diseases and Medical Procedures. TH Open. 2022 Jul

Subcellular Location

Membrane.

UNIPROT

SwissProt: P07204 Human

Synonyms

AHUS 6 antibody

AHUS6 antibody

BDCA 3 antibody

BDCA3 antibody

CD 141 antibody

CD141 antibody

CD141 antigen antibody

Fetomodulin antibody

Thbd antibody

THPH12 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Thrombomodulin on different lysates with Rabbit anti-Thrombomodulin antibody (<a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a>) at 1/20,000 dilution. <br /> <br />Lane 1: A431 (Human epidermoid carcinoma skin squamous cells) cell lysate <br />Lane 2: 293T (Human embryonic kidney cells) cell lysate <br />Lane 3: THP-1 (Human acute monoblastic leukemia cells) cell lysate <br />Lane 4: Jurkat (Human T-lymphoblastic cells) cell lysate <br /> <br />Lysates/proteins at 10 µg/Lane. <br />Exposure time: 2 minutes; ECL: K1801 <br /> <br />293T is negative control (PMID: 40414460). <br />Negative expression of Thrombomodulin protein in Jurkat cell is consistent with the predicted expression pattern. <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a>, 1/20,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 60 kDa <br />Observed band size: 75-100 kDa

    ☑ Relative expression (RE)

    Western blot analysis of Thrombomodulin on different lysates with Rabbit anti-Thrombomodulin antibody (HA751888) 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: HA751888, 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

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Thrombomodulin on different lysates with Rabbit anti-Thrombomodulin antibody (<a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a>) at 1/5,000 dilution. <br /> <br />Lane 1: A431 (Human epidermoid carcinoma skin squamous cells) cell lysate <br />Lane 2: A431 treated with PNGase F cell lysate <br /> <br />Lysates/proteins at 10 µg/Lane. <br />Exposure time: 1 minute; ECL: K1801 <br /> <br />Thrombomodulin is a glycoprotein and can be deglycosylated by PNGase F. The molecular mass observed is consistent with the literature (PMID: 2827310) <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 60 kDa <br />Observed band size: 75-100 kDa

    ☑ Cell treatment (CT)

    Western blot analysis of Thrombomodulin on different lysates with Rabbit anti-Thrombomodulin antibody (HA751888) 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: HA751888, 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

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: Immunocytochemistry (IF-cell) <br /> <br />Species: Human <br />Sample: A431 (Human epidermoid carcinoma skin squamous cells)/293T (Human embryonic kidney cells) <br /> <br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. <br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature. <br /> <br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature. <br />Antibody dilution buffer: 1% BSA in PBST. <br />Primary antibody: <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a>, 1/500, overnight at 4°C. <br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature. <br /> <br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue). <br />Negative control: 293T (PMID: 40414460).

    ☑ Relative expression (RE)

    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: HA751888, 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).

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Human <br />Tissue: Placenta <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a>, 1/1,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    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: HA751888, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: squamous cell lung carcinoma<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a>, 1/200, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    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: HA751888, 1/200, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Human <br />Tissue: Lymph node <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a>, 1/200, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    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: HA751888, 1/200, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Esophagus <br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a>, 1/200, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    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: HA751888, 1/200, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Ovarian carcinoma<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a>, 1/200, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    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: HA751888, 1/200, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: Flow Cytometry <br /> <br />Species: Human <br />Sample: A431 (Human epidermoid carcinoma skin squamous cells)/293T (Human embryonic kidney cells) <br /> <br />Blocking: 1% BSA + 10% normal goat serum + human FcR (Miltenyi Biotec), 15 minutes at room temperature. (human) <br />Antibody dilution buffer: 1x PBS. <br />Primary antibody: <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a> (1/10,000) (Red) compared with Rabbit IgG Isotype Control (<a href="/products/HA722127" style="font-weight: bold;text-decoration: underline;">HA722127</a>, Green), 15 minutes at room temperature. <br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 15 minutes at room temperature. <br />Negative control: 293T (PMID: 40414460).

    ☑ Relative expression (RE)

    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: HA751888 (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).

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: Flow Cytometry <br /> <br />Species: Human <br />Sample: A431 (Human epidermoid carcinoma skin squamous cells)/Jurkat (Human T-lymphoblastic cells) <br /> <br />Blocking: 1% BSA + 10% normal goat serum + human FcR (Miltenyi Biotec), 15 minutes at room temperature. (human) <br />Antibody dilution buffer: 1x PBS. <br />Primary antibody: <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a> (1/10,000) (Red) compared with Rabbit IgG Isotype Control (<a href="/products/HA722127" style="font-weight: bold;text-decoration: underline;">HA722127</a>, Green), 15 minutes at room temperature. <br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 15 minutes at room temperature. <br />Negative expression of Thrombomodulin protein in Jurkat cell is consistent with the predicted expression pattern.

    ☑ Relative expression (RE)

    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: HA751888 (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.

  • Immunoprecipitation (IP)<br /><br />Thrombomodulin was immunoprecipitated in 0.2 mg THP-1 (Human acute monoblastic leukemia cells) cell lysate with <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a> 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.<br /><br />Lane 1: THP-1 cell lysate (input)<br />Lane 2: <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a> IP in THP-1 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a> in THP-1 cell lysate<br /><br />Exposure time: 8 seconds<br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br /><br />Primary dilution: <a href="/products/HA751888" style="font-weight: bold;text-decoration: underline;">HA751888</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), 2 hours at room temperature<br /><br />Predicted band size: 60 kDa<br />Observed band size: 75-100 kDa

    Immunoprecipitation (IP)

    Thrombomodulin was immunoprecipitated in 0.2 mg THP-1 (Human acute monoblastic leukemia cells) cell lysate with HA751888 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751888 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: HA751888 IP in THP-1 cell lysate
    Lane 3: Rabbit IgG instead of HA751888 in THP-1 cell lysate

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

    Primary dilution: HA751888, 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

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"