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CD86 Recombinant Rabbit Monoclonal Antibody [SJ20-00]

Usd: 205

Specification

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Catalog# ET1606-50

CD86 Recombinant Rabbit Monoclonal Antibody [SJ20-00]

Application

  • WB

  • IF-Cell

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

CD86 Recombinant Rabbit Monoclonal Antibody [SJ20-00]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human CD86 aa 1-50 / 329.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IP

Molecular Weight

Predicted band size: 38 kDa

Positive Control

Daudi cell lysate, Raji cell lysate, Ramos cell lysate, K-562 cell lysate, Mouse lung tissue lysate, Rat lung tissue lysate, Raji.

Conjugation

unconjugated

Clone Number

SJ20-00

RRID

AB_3069745

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000-1:20,000

  • IF-Cell

  • 1:500

  • IP

  • Use at an assay dependent concentration.

Target

Function

Cluster of Differentiation 86 (also known as CD86 and B7-2) is a protein constitutively expressed on dendritic cells, Langerhans cells, macrophages, B-cells (including memory B-cells), and on other antigen-presenting cells. Along with CD80, CD86 provides costimulatory signals necessary for T cell activation and survival. Depending on the ligand bound, CD86 can signal for self-regulation and cell-cell association, or for attenuation of regulation and cell-cell disassociation. The CD86 gene encodes a type I membrane protein that is a member of the immunoglobulin superfamily. Alternative splicing results in two transcript variants encoding different isoforms. Additional transcript variants have been described, but their full-length sequences have not been determined. Roles of both CD80 and CD86 are studied in context of many pathologies. Selective inhibition of costimulatory inhibitors was examined in a model of allergic pulmonary inflammation and airway hyper-responsiveness (AHR). Since initial host response to Staphylococcus aureus, especially the immune response based on T cells, is a contributing factor in the pathogenesis of acute pneumonia, role of the CD80/CD86 pathway in pathogenesis was investigated. The costimulatory molecules were also investigated in context of Bronchial Astma, Treg in cancer, and immunotherapy.

Background References

1. Li Y & Ding J Optimized generation of survivin-specific cytotoxic T lymphocytes against lung cancer. Mol Med Rep 12:2169-74 (2015).

2. Tarhini AA et al. Immune monitoring of the circulation and the tumor microenvironment in patients with regionally advanced melanoma receiving neoadjuvant ipilimumab. PLoS One 9:e87705 (2014).

Tissue Specificity

Expressed by activated B-lymphocytes and monocytes.

Post-translational Modification

Polyubiquitinated; which is promoted by MARCH8 and results in endocytosis and lysosomal degradation.

Subcellular Location

Cell membrane.

UNIPROT

SwissProt: P42081 Human

SwissProt: P42082 Mouse

Entrez Gene: 56822 Rat

Synonyms

Activation B7-2 antigen 3 antibody

Activation B7-2 antigen antibody

B-lymphocyte activation antigen B7-2 2 antibody

B-lymphocyte activation antigen B7-2 antibody

B7 2 antibody

B7 antibody

B7-2 antibody

B7.2 antibody

B70 antibody

B72 antigen antibody

Expand

Images

  • Western blot analysis of CD86 on different lysates with Rabbit anti-CD86 antibody (<a href="/products/ET1606-50" style="font-weight: bold;text-decoration: underline;">ET1606-50</a>) at 1/5,000 dilution.<br /><br />Lane 1: Daudi cell lysate<br />Lane 2: Raji cell lysate<br />Lane 3: Ramos cell lysate<br />Lane 4: K-562 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 38 kDa<br />Observed band size: 70 kDa<br /><br />Exposure time: 2 minutes 37 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/ET1606-50" style="font-weight: bold;text-decoration: underline;">ET1606-50</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. 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.

    Western blot analysis of CD86 on different lysates with Rabbit anti-CD86 antibody (ET1606-50) at 1/5,000 dilution.

    Lane 1: Daudi cell lysate
    Lane 2: Raji cell lysate
    Lane 3: Ramos cell lysate
    Lane 4: K-562 cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 38 kDa
    Observed band size: 70 kDa

    Exposure time: 2 minutes 37 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 (ET1606-50) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of CD86 on different lysates with Rabbit anti-CD86 antibody (<a href="/products/ET1606-50" style="font-weight: bold;text-decoration: underline;">ET1606-50</a>) at 1/5,000 dilution.<br /><br />Lane 1: Raji cell lysate (10 µg/Lane)<br />Lane 2: Mouse lung tissue lysate (20 µg/Lane)<br />Lane 3: Rat lung tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 38 kDa<br />Observed band size: 70 kDa<br /><br />Exposure time: 1 minute;<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/ET1606-50" style="font-weight: bold;text-decoration: underline;">ET1606-50</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. 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.

    Western blot analysis of CD86 on different lysates with Rabbit anti-CD86 antibody (ET1606-50) at 1/5,000 dilution.

    Lane 1: Raji cell lysate (10 µg/Lane)
    Lane 2: Mouse lung tissue lysate (20 µg/Lane)
    Lane 3: Rat lung tissue lysate (20 µg/Lane)

    Predicted band size: 38 kDa
    Observed band size: 70 kDa

    Exposure time: 1 minute;

    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 (ET1606-50) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of Raji cells labeling CD86 with Rabbit anti-CD86 antibody (<a href="/products/ET1606-50" style="font-weight: bold;text-decoration: underline;">ET1606-50</a>) at 1/500 dilution.<br /><br />Cells were fixed in 80% precooled methanol for 5 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-CD86 antibody (<a href="/products/ET1606-50" style="font-weight: bold;text-decoration: underline;">ET1606-50</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 Raji cells labeling CD86 with Rabbit anti-CD86 antibody (ET1606-50) at 1/500 dilution.

    Cells were fixed in 80% precooled methanol for 5 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-CD86 antibody (ET1606-50) 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.

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

Citation

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