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CD63 Recombinant Rabbit Monoclonal Antibody [SY21-02] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750112

CD63 Recombinant Rabbit Monoclonal Antibody [SY21-02] - BSA and Azide free

Application

  • WB

  • IHC-P

Reactivity

  • Human

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

CD63 Recombinant Rabbit Monoclonal Antibody [SY21-02] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human CD63 aa 88-216 / 238.

Species Reactivity

Human

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 26 kDa

Positive Control

SK-MEL-28 cell lysate, U-87 MG cell lysate, human lung carcinoma tissue, human tonsil tissue, human spleen tissue.

Conjugation

unconjugated

Clone Number

SY21-02

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:100-1:500

Target

Function

The tetraspanins are integral membrane proteins expressed on cell surface and granular membranes of hematopoietic cells and are components of multi-molecular complexes with specific integrins. The tetraspanin CD63 (also known as LAMP-3, Melanoma-associated antigen ME491, TSPAN30, MLA1 and OMA81H) is a lysosomal membrane glycoprotein that translocates to the plasma membrane after platelet activation. CD63 is expressed on activated platelets, monocytes and macrophages, and is weakly expressed on granulocytes, T cell and B cells. It is located on the basophilic granule membranes and on the plasma membranes of lymphocytes and granulocytes. CD63 is a member of the TM4 superfamily of leukocyte glycoproteins that includes CD9, CD37 and CD53, which contain four transmembrane regions. CD63 may play a role in phagocytic and intracellular lysosome-phagosome fusion events. CD63 deficiency is associated with Hermansky-Pudlak syndrome.

Background References

1. Ghossoub R et al. Syntenin-ALIX exosome biogenesis and budding into multivesicular bodies are controlled by ARF6 and PLD2. Nat Commun 5:3477 (2014).

2. Rodríguez M et al. Different exosome cargo from plasma/bronchoalveolar lavage in non-small-cell lung cancer. Genes Chromosomes Cancer 53:713-24 (2014).

Sequence Similarity

Belongs to the tetraspanin (TM4SF) family.

Tissue Specificity

Detected in platelets (at protein level). Dysplastic nevi, radial growth phase primary melanomas, hematopoietic cells, tissue macrophages.

Post-translational Modification

Palmitoylated at a low, basal level in unstimulated platelets. The level of palmitoylation increases when platelets are activated by thrombin (in vitro).

Subcellular Location

Lysosome membrane, Cell membrane, extracellular exosome, Late endosome membrane, multivesicular body, Melanosome, Cell surface.

UNIPROT

SwissProt: P08962 Human

Synonyms

Lysosomal associated membrane protein 3 antibody

CD 63 antibody

CD63 antibody

CD63 antigen (melanoma 1 antigen) antibody

CD63 antigen antibody

CD63 antigen melanoma 1 antigen antibody

CD63 molecule antibody

CD63_HUMAN antibody

gp55 antibody

Granulophysin antibody

Expand

Images

  • Western blot analysis of CD63 on different lysates with Rabbit anti-CD63 antibody (<a href="/products/HA750112" style="font-weight: bold;text-decoration: underline;">HA750112</a>) at 1/2,000 dilution.<br /><br />Lane 1: SK-MEL-28 cell lysate<br />Lane 2: U-87 MG cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 26 kDa<br />Observed band size: 30~60 kDa<br /><br />Exposure time: 43 seconds;<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/HA750112" style="font-weight: bold;text-decoration: underline;">HA750112</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.

    Western blot analysis of CD63 on different lysates with Rabbit anti-CD63 antibody (HA750112) at 1/2,000 dilution.

    Lane 1: SK-MEL-28 cell lysate
    Lane 2: U-87 MG cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 26 kDa
    Observed band size: 30~60 kDa

    Exposure time: 43 seconds;

    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 (HA750112) 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.

  • Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-CD63 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750112" style="font-weight: bold;text-decoration: underline;">HA750112</a>, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-CD63 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750112, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CD63 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750112" style="font-weight: bold;text-decoration: underline;">HA750112</a>, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CD63 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750112, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-CD63 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750112" style="font-weight: bold;text-decoration: underline;">HA750112</a>, 1/400) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-CD63 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750112, 1/400) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of CD63 on different lysates with Rabbit anti-CD63 antibody (<a href="/products/HA750112" style="font-weight: bold;text-decoration: underline;">HA750112</a>) at 1/2,000 dilution.<br /><br />Lane 1: SK-MEL-28-si NT cell lysate<br />Lane 2: SK-MEL-28-si CD63 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 26 kDa<br />Observed band size: 30~60 kDa<br /><br />Exposure time: 1 minute 30 seconds;<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/HA750112" style="font-weight: bold;text-decoration: underline;">HA750112</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 1/100,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of CD63 on different lysates with Rabbit anti-CD63 antibody (HA750112) at 1/2,000 dilution.

    Lane 1: SK-MEL-28-si NT cell lysate
    Lane 2: SK-MEL-28-si CD63 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 26 kDa
    Observed band size: 30~60 kDa

    Exposure time: 1 minute 30 seconds;

    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 (HA750112) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

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

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