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CD27 Recombinant Rabbit Monoclonal Antibody [PSH24-12]

Usd: 385

Specification

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Catalog# HA724423

CD27 Recombinant Rabbit Monoclonal Antibody [PSH24-12]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC(Intra)

Reactivity

  • Human

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

CD27 Recombinant Rabbit Monoclonal Antibody [PSH24-12]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IHC-P, FC(Intra)

Molecular Weight

Predicted band size: 29 kDa

Conjugation

unconjugated

Clone Number

PSH24-12

Product Features

Form

Liquid

Concentration

Storage Instructions

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.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:10,000

  • IF-Cell

  • 1:2,000

  • IHC-P

  • 1:500

  • FC(Intra)

  • 1:1,000

Target

Function

The tumor necrosis factor (TNF) receptor family is composed of several type I integral membrane glycoproteins that exhibit homology in their cysteine-rich extracellular domains. Members of this family include FAS, OX40, CD27 and CD30. Ligands for these receptors are often type II transmembrane glycoproteins, as is the case for CD27 and CD30. CD27 is a homodimeric lymphocyte-specific surface antigen present on T and B lymphocytes. Activation of the CD3 complex via the T cell receptor for antigen leads to an increase in CD27 expression. Together, CD27 and its ligand, CD27L, generate co-stimulatory signals required for complete T cell activation. CD30 is a surface marker for neoplastic cells of the Hodgkin's lymphoma and related hematologic malignancies. CD30L has been shown to enhance the proliferation of the Hodgkin's cell line HDLM-2, but exerts antiproliferative effects on large cell anaplastic lymphoma cell lines.

Background References

1. Grimsholm O. CD27 on human memory B cells-more than just a surface marker. Clin Exp Immunol. 2023 Jul

2. Starzer AM et al. New emerging targets in cancer immunotherapy: CD27 (TNFRSF7). ESMO Open. 2020 Mar

Subcellular Location

Cell membrane.

UNIPROT

SwissProt: P26842 Human

Synonyms

CD 27 antibody

CD27 antibody

CD27 antigen antibody

CD27 molecule antibody

CD27_HUMAN antibody

CD27L receptor antibody

LPFS2 antibody

MGC20393 antibody

OTTHUMP00000238557 antibody

S152 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of CD27 on different lysates with Rabbit anti-CD27 antibody (<a href="/products/HA724423" style="font-weight: bold;text-decoration: underline;">HA724423</a>) at 1/10,000 dilution.<br /><br />Lane 1: Raji (Human Burkitt's lymphoma cells) cell lysate<br />Lane 2: MCF7 (Human breast cancer cells) cell lysate<br />Lane 3: Ramos (Human Burkitt's lymphoma cells) cell lysate<br />Lane 4: Daudi (Human Burkitt's lymphoma cells) cell lysate<br />Lane 5: THP-1 (Human acute monoblastic leukemia cells) cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br />Exposure time: 12 seconds; ECL: K1801<br /><br />MCF7 and THP-1 are negative controls (PMID: 11062504).<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA724423" style="font-weight: bold;text-decoration: underline;">HA724423</a>, 1/10,000 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: 29 kDa<br />Observed band size: 50 kDa

    ☑ Relative expression (RE)

    Western blot analysis of CD27 on different lysates with Rabbit anti-CD27 antibody (HA724423) at 1/10,000 dilution.

    Lane 1: Raji (Human Burkitt's lymphoma cells) cell lysate
    Lane 2: MCF7 (Human breast cancer cells) cell lysate
    Lane 3: Ramos (Human Burkitt's lymphoma cells) cell lysate
    Lane 4: Daudi (Human Burkitt's lymphoma cells) cell lysate
    Lane 5: THP-1 (Human acute monoblastic leukemia cells) cell lysate

    Lysates/proteins at 10 µg/Lane.
    Exposure time: 12 seconds; ECL: K1801

    MCF7 and THP-1 are negative controls (PMID: 11062504).

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724423, 1/10,000 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: 29 kDa
    Observed band size: 50 kDa

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: Immunocytochemistry (IF-cell) <br /> <br />Species: Human <br />Sample: Raji (Human Burkitt's lymphoma cells) / MCF7 (Human breast cancer cells) <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/HA724423" style="font-weight: bold;text-decoration: underline;">HA724423</a>, 1/2,000, 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: MCF7 (PMID: 11062504).

    ☑ Relative expression (RE)

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: Raji (Human Burkitt's lymphoma cells) / MCF7 (Human breast cancer 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: HA724423, 1/2,000, 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: MCF7 (PMID: 11062504).

  • 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°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/HA724423" style="font-weight: bold;text-decoration: underline;">HA724423</a>, 1/500, 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°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: HA724423, 1/500, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Human <br />Tissue: Lymph nodes <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/HA724423" style="font-weight: bold;text-decoration: underline;">HA724423</a>, 1/500, 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 nodes
    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: HA724423, 1/500, 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: Immunohistochemistry (IHC-P) <br /> <br />Species: Human <br />Tissue: Skeletal muscle <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/HA724423" style="font-weight: bold;text-decoration: underline;">HA724423</a>, 1/500, 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. <br /> <br />Negative expression of CD27 in human skeletal muscle is consistent with the predicted expression pattern.

    ☑ Relative expression (RE)

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Skeletal muscle
    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: HA724423, 1/500, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

    Negative expression of CD27 in human skeletal muscle is consistent with the predicted expression pattern.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: Flow Cytometry (Intra) <br /> <br />Species: Human <br />Sample: Raji (Human Burkitt's lymphoma cells) / MCF7 (Human breast cancer cells) <br /> <br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. <br />Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature. <br /> <br />Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature. <br />Antibody dilution buffer: 1x PBS. <br />Primary antibody: <a href="/products/HA724423" style="font-weight: bold;text-decoration: underline;">HA724423</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. <br /> <br />Negative control: MCF7 (PMID: 11062504).

    ☑ Relative expression (RE)

    Application: Flow Cytometry (Intra)

    Species: Human
    Sample: Raji (Human Burkitt's lymphoma cells) / MCF7 (Human breast cancer cells)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.

    Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
    Antibody dilution buffer: 1x PBS.
    Primary antibody: HA724423 (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.

    Negative control: MCF7 (PMID: 11062504).

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

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