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CD24 Recombinant Rabbit Monoclonal Antibody [PSH10-85] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751377

CD24 Recombinant Rabbit Monoclonal Antibody [PSH10-85] - BSA and Azide free

Application

  • IF-Cell

  • FC

  • WB

Reactivity

  • Human

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

CD24 Recombinant Rabbit Monoclonal Antibody [PSH10-85] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human CD24 aa

Species Reactivity

Human

Validated Applications

IF-Cell, FC, WB

Molecular Weight

Predicted band size: 8 kDa

Positive Control

MCF7, MCF7 cell lysate, HT-29 cell lysate.

Conjugation

unconjugated

Clone Number

PSH10-85

Product Features

Form

Liquid

Concentration

1mg/ml

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

  • IF-Cell

  • 1:500

  • WB

  • 1:2,000-1:10,000

  • FC

  • 1:1,000

Target

Function

Signal transducer CD24 also known as cluster of differentiation 24 or heat stable antigen CD24 (HSA) is a protein that in humans is encoded by the CD24 gene. CD24 is a cell adhesion molecule. CD24 is a sialoglycoprotein expressed at the surface of most B lymphocytes and differentiating neuroblasts. It is also expressed on neutrophils and neutrophil precursors from the myelocyte stage onwards. The encoded protein is anchored via a glycosyl phosphatidylinositol (GPI) link to the cell surface. The protein also contributes to a wide range of downstream signaling networks and is crucial for neural development. Cross-linking of CD24 on the surface of neutrophils induces apoptosis, and this appears to be defective in sepsis.

Background References

1. Sun F et al. Bispecific BCMA/CD24 CAR-T cells control multiple myeloma growth. Nat Commun. 2024 Jan

2. Liu Y et al. CD24-Siglec interactions in inflammatory diseases. Front Immunol. 2023 May

Subcellular Location

Cell membrane.

UNIPROT

SwissProt: P25063 Human

Synonyms

CD 24 antibody

CD24 antibody

CD24 antigen (small cell lung carcinoma cluster 4 antigen) antibody

CD24 antigen antibody

CD24 molecule antibody

CD24_HUMAN antibody

CD24A antibody

FLJ22950 antibody

FLJ43543 antibody

GPI linked surface mucin antibody

Expand

Images

  • Immunocytochemistry analysis of MCF7 cells labeling CD24 with Rabbit anti-CD24 antibody (<a href="/products/HA751377" style="font-weight: bold;text-decoration: underline;">HA751377</a>) at 1/500 dilution.<br /><br />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-CD24 antibody (<a href="/products/HA751377" style="font-weight: bold;text-decoration: underline;">HA751377</a>) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of MCF7 cells labeling CD24 with Rabbit anti-CD24 antibody (HA751377) 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-CD24 antibody (HA751377) 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of CD24 on different lysates with Rabbit anti-CD24 antibody (<a href="/products/HA751377" style="font-weight: bold;text-decoration: underline;">HA751377</a>) at 1/2,000 dilution.<br /><br />Lane 1: MCF7 cell lysate<br />Lane 2: MCF7 cell lysate treated with deglycosylation<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 8 kDa<br />Observed band size: 20-55 kDa<br /><br />Exposure time: 14 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA751377" style="font-weight: bold;text-decoration: underline;">HA751377</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of CD24 on different lysates with Rabbit anti-CD24 antibody (HA751377) at 1/2,000 dilution.

    Lane 1: MCF7 cell lysate
    Lane 2: MCF7 cell lysate treated with deglycosylation

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 8 kDa
    Observed band size: 20-55 kDa

    Exposure time: 14 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 (HA751377) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of CD24 on different lysates with Rabbit anti-CD24 antibody (<a href="/products/HA751377" style="font-weight: bold;text-decoration: underline;">HA751377</a>) at 1/5,000 dilution.<br /><br />Lane 1: MCF7 cell lysate<br />Lane 2: MDA-MB-231 cell lysate (negative)<br />Lane 3: HCT 116 cell lysate (negative)<br />Lane 4: HT-29 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 8 kDa<br />Observed band size: 25-40 kDa<br /><br />Exposure time: 4 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA751377" style="font-weight: bold;text-decoration: underline;">HA751377</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of CD24 on different lysates with Rabbit anti-CD24 antibody (HA751377) at 1/5,000 dilution.

    Lane 1: MCF7 cell lysate
    Lane 2: MDA-MB-231 cell lysate (negative)
    Lane 3: HCT 116 cell lysate (negative)
    Lane 4: HT-29 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 8 kDa
    Observed band size: 25-40 kDa

    Exposure time: 4 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 (HA751377) 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.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Flow cytometric analysis of MDA-MB-231 (left, negative) and MCF7 (right, positive) cells labeling CD24.<br /><br />Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (<a href="/products/HA751377" style="font-weight: bold;text-decoration: underline;">HA751377</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    ☑ Relative expression (RE)

    Flow cytometric analysis of MDA-MB-231 (left, negative) and MCF7 (right, positive) cells labeling CD24.

    Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA751377, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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