PRODUCT CODE: ET1606-50

CD86 Recombinant Rabbit Monoclonal Antibody [SJ20-00] (ET1606-50)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Rat

  • Monkey

"Western blot analysis of CD86 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1606-50, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Raji cell lysate<br />
Lane 2: Hela cell lysate<br />
Lane 3: HepG2 cell lysate<br />
Lane 4: MCF-7 cell lysate
  • "Western blot analysis of CD86 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1606-50, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Raji cell lysate<br />
Lane 2: Hela cell lysate<br />
Lane 3: HepG2 cell lysate<br />
Lane 4: MCF-7 cell lysate
  • ICC staining of CD86 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1606-50, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of CD86 in HUVEC cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1606-50, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of CD86 in JAR cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1606-50, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CD86 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 (ET1606-50, 1/50) 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.
  • Flow cytometric analysis of CD86 was done on K562 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1606-50, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
  • Flow cytometric analysis of CD86 was done on Raji cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1606-50, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
"Western blot analysis of CD86 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1606-50, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Raji cell lysate
Lane 2: Hela cell lysate
Lane 3: HepG2 cell lysate
Lane 4: MCF-7 cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Rat

  • Monkey

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

CD86 Recombinant Rabbit Monoclonal Antibody [SJ20-00] (ET1606-50)

Immunogen

Synthetic peptide within human cd86 aa 1-50 / 329.

Host

Rabbit

Positive Control

Hela, HUVEC, JAR, human tonsil tissue, K562, Raji.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SJ20-00

PROPERTIES

Form

Liquid

Storage Condition

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

Storage Buffer

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

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

70 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

CD86

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; BU63 antibody; CD28 antigen ligand 2 2 antibody; CD28 antigen ligand 2 antibody; Cd28l2 antibody; CD28LG2 antibody; CD86 antibody; CD86 antigen (CD28 antigen ligand 2, B7-2 antigen) 1, 2 antibody; CD86 antigen (CD28 antigen ligand 2, B7-2 antigen) antibody; CD86 antigen antibody; CD86 molecule antibody; CD86_HUMAN antibody; CLS1 antibody; CTLA-4 counter-receptor B7.2 2, 3 antibody; CTLA-4 counter-receptor B7.2 antibody; Early T-cell costimulatory molecule 1 antibody; ETC-1 antibody; FUN 1 antibody; FUN-1 antibody; LAB72 antibody; Ly-58 antibody; Ly58 antibody; MB7 antibody; MB7-2 antibody; Membrane glycoprotein antibody; MGC34413 antibody; T-lymphocyte activation antigen CD86 antibody; TS/A-2 antibody

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.

FUNCTION

T cell proliferation and lymphokine production are triggered by occupation of the TCR by antigen, followed by a costimulatory signal that is delivered by a ligand expressed on antigen presenting cells. The B7-related cell surface proteins B7-1 (CD80) and B7-2 (CD86) expressed on antigen presenting cells bind the homologous T cell receptors CD28 and CTLA-4 (cytotoxic T lymphocyte-associated protein-4) and trigger costimulatory signals for optimal T cell activation. CTLA-4 shares 31% overall amino acid identity with CD28, and it has been proposed that CD28 and CTLA-4 are functionally redundant. SLAM is a novel receptor on T cells that, when engaged, potentiates T cell expansion in a CD28-independent manner. B7, also designated BB1, is another ligand or counterreceptor for CD28 and CTLA-4 that is expressed on the antigen-presenting cell.