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ICAM3 Recombinant Rabbit Monoclonal Antibody [PSH20-97] - BSA and Azide free

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Catalog# HA751779

ICAM3 Recombinant Rabbit Monoclonal Antibody [PSH20-97] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

  • IP

Reactivity

  • Human

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

ICAM3 Recombinant Rabbit Monoclonal Antibody [PSH20-97] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human ICAM3 aa 1-485.

Species Reactivity

Human

Validated Applications

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

Molecular Weight

Predicted band size: 60 kDa

Positive Control

U-937 (Human acute monocytic leukemia cells) cell lysate, Jurkat (Human T-lymphoblastic cells) cell lysate, Ramos (Human Burkitt's lymphoma cells) cell lysate, THP-1 (Human acute monoblastic leukemia cells) cell lysate, HL-60 (Human acute promyelocytic leukemia cells) cell lysate.

Conjugation

unconjugated

Clone Number

PSH20-97

Product Features

Form

Liquid

Concentration

Storage Instructions

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

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IF-Cell

  • 1:500

  • IHC-P

  • 1:1,000

  • FC

  • 1:1,000

  • IP

  • 1-2μg/sample

Target

Function

Intercellular adhesion molecule 3 (ICAM3) also known as CD50 (Cluster of Differentiation 50), is a protein that in humans is encoded by the ICAM3 gene. The protein is constitutively expressed on the surface of leukocytes, which are also called white blood cells and are part of the immune system. ICAM3 mediates adhesion between cells by binding to specific integrin receptors. It plays an important role in the immune cell response through its facilitation of interactions between T cells and dendritic cells, which allows for T cell activation. ICAM3 also mediates the clearance of cells undergoing apoptosis by attracting and binding macrophages, a type of cell that breaks down infected or dying cells through a process known as phagocytosis, to apoptotic cells.

Background References

1. Li L et al. Functional evaluation of various ICAM3 transcript variants in diffuse large B-Cell lymphoma. Leuk Lymphoma. 2022 Dec

2. Zicheng H et al. Association of Circulating ICAM3 Concentrations with Severity and Short-term Outcomes of Acute Ischemic Stroke. Neurotox Res. 2021 Aug

Subcellular Location

Membrane.

UNIPROT

SwissProt: P32942 Human

Synonyms

CD50 antibody

CDW 50 antibody

CDw50 antibody

ICAM 3 antibody

ICAM-3 antibody

ICAM-R antibody

ICAM3 antibody

ICAM3_HUMAN antibody

ICAMR antibody

Intercellular adhesion molecule 3 antibody

Images

  • Western blot analysis of ICAM3 on different lysates with Rabbit anti-ICAM3 antibody (<a href="/products/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</a>) at 1/5,000 dilution.<br /><br />Lane 1: U-937 (Human acute monocytic leukemia cells) cell lysate<br />Lane 2: Jurkat (Human T-lymphoblastic cells) cell lysate<br />Lane 3: Ramos (Human Burkitt's lymphoma cells) cell lysate<br />Lane 4: THP-1 (Human acute monoblastic leukemia cells) cell lysate<br />Lane 5: HL-60 (Human acute promyelocytic leukemia cells) cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br />Exposure time: 20 seconds; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</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 ℃<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: 100-150 kDa

    Western blot analysis of ICAM3 on different lysates with Rabbit anti-ICAM3 antibody (HA751779) at 1/5,000 dilution.

    Lane 1: U-937 (Human acute monocytic leukemia cells) cell lysate
    Lane 2: Jurkat (Human T-lymphoblastic cells) cell lysate
    Lane 3: Ramos (Human Burkitt's lymphoma cells) cell lysate
    Lane 4: THP-1 (Human acute monoblastic leukemia cells) cell lysate
    Lane 5: HL-60 (Human acute promyelocytic leukemia cells) cell lysate

    Lysates/proteins at 15 µg/Lane.
    Exposure time: 20 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA751779, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    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: 100-150 kDa

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of ICAM3 on different lysates with Rabbit anti-ICAM3 antibody (<a href="/products/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</a>) at 1/5,000 dilution.<br /><br />Lane 1: Jurkat (Human T-lymphoblastic cells) cell lysate<br />Lane 2: Jurkat treated with PNGseF cell lysate<br />Lane 3: U-937 (Human acute monocytic leukemia cells) cell lysate<br />Lane 4: U-937 treated with PNGseF cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br />Exposure time: 3 minutes; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</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 ℃<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: 100-150/60 kDa

    ☑ Cell treatment (CT)

    Western blot analysis of ICAM3 on different lysates with Rabbit anti-ICAM3 antibody (HA751779) at 1/5,000 dilution.

    Lane 1: Jurkat (Human T-lymphoblastic cells) cell lysate
    Lane 2: Jurkat treated with PNGseF cell lysate
    Lane 3: U-937 (Human acute monocytic leukemia cells) cell lysate
    Lane 4: U-937 treated with PNGseF cell lysate

    Lysates/proteins at 10 µg/Lane.
    Exposure time: 3 minutes; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA751779, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    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: 100-150/60 kDa

  • Application: Immunocytochemistry (IF-cell)<br /><br />Species: Human<br />Sample: Jurkat (Human T-lymphoblastic 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/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</a>, 1/500, overnight at 4℃.<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).

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: Jurkat (Human T-lymphoblastic 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: HA751779, 1/500, overnight at 4℃.
    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).

  • Application: Immunocytochemistry (IF-cell)<br /><br />Species: Human<br />Sample: U-937 (Human acute monocytic leukemia 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/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</a>, 1/500, overnight at 4℃.<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).

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: U-937 (Human acute monocytic leukemia 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: HA751779, 1/500, overnight at 4℃.
    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).

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Tonsil<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/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</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: Tonsil
    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: HA751779, 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: Colon<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/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</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: Colon
    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: HA751779, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Flow Cytometry<br /><br />Species: Human<br />Sample: 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/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</a> (1/1,000) (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.

    Application: Flow Cytometry

    Species: Human
    Sample: 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: HA751779 (1/1,000) (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.

  • Application: Flow Cytometry<br /><br />Species: Human<br />Sample: U-937 (Human acute monocytic leukemia 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/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</a> (1/1,000) (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.

    Application: Flow Cytometry

    Species: Human
    Sample: U-937 (Human acute monocytic leukemia 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: HA751779 (1/1,000) (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.

  • Immunoprecipitation (IP)<br /><br />ICAM3 was immunoprecipitated in 0.2 mg U-937 (Human acute monocytic leukemia cells) cell lysate with <a href="/products/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</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: U-937 cell lysate (input)<br />Lane 2: <a href="/products/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</a> IP in U-937 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</a> in U-937 cell lysate<br /><br />Exposure time: 14 seconds<br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br /><br />Primary dilution: <a href="/products/HA751779" style="font-weight: bold;text-decoration: underline;">HA751779</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: 100-150 kDa

    Immunoprecipitation (IP)

    ICAM3 was immunoprecipitated in 0.2 mg U-937 (Human acute monocytic leukemia cells) cell lysate with HA751779 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751779 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: U-937 cell lysate (input)
    Lane 2: HA751779 IP in U-937 cell lysate
    Lane 3: Rabbit IgG instead of HA751779 in U-937 cell lysate

    Exposure time: 14 seconds
    Blocking: 5% NFDM/TBST, 1 hour at room temperature

    Primary dilution: HA751779, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

    Predicted band size: 60 kDa
    Observed band size: 100-150 kDa

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

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