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FYB Recombinant Rabbit Monoclonal Antibody [JE44-80]

Usd: 385

Specification

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Catalog# ET7109-38

FYB Recombinant Rabbit Monoclonal Antibody [JE44-80]

Application

  • WB

  • IHC-P

  • FC

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

FYB Recombinant Rabbit Monoclonal Antibody [JE44-80]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human FYB aa 701-750 / 783.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, FC, IF-Cell

Molecular Weight

Predicted band size: 85 kDa

Positive Control

U-937 cell lysate, Jurkat cell lysate, RAW264.7 cell lysate, mouse spleen tissue lysate, human tonsil tissue, human appendix tissue, human spleen tissue, rat spleen tissue, HepG2.

Conjugation

unconjugated

Clone Number

JE44-80

RRID

AB_3070925

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

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

  • IF-Cell

  • 1:100

Target

Function

Acts as an adapter protein of the FYN and LCP2 signaling cascades in T-cells. Modulates the expression of interleukin-2 (IL-2). Involved in platelet activation. Prevents the degradation of SKAP1 and SKAP2. May play a role in linking T-cell signaling to remodeling of the actin cytoskeleton. Expressed in hematopoietic tissues such as myeloid and T-cells, spleen and thymus. Not expressed in B-cells, nor in non-lymphoid tissues.

Background References

1. Krause M et al. Fyn-binding protein (Fyb)/SLP-76-associated protein (SLAP), Ena/vasodilator-stimulated phosphoprotein (VASP) proteins and the Arp2/3 complex link T cell receptor (TCR) signaling to the actin cytoskeleton. J Cell Biol 149:181-194 (2000).

2. Huang Y et al. Deficiency of ADAP/Fyb/SLAP-130 destabilizes SKAP55 in Jurkat T cells. J Biol Chem 280:23576-23583 (2005).

Tissue Specificity

Expressed in hematopoietic tissues such as myeloid and T-cells, spleen and thymus. Not expressed in B-cells, nor in non-lymphoid tissues.

Post-translational Modification

T-cell receptor ligation leads to increased tyrosine phosphorylation.

Subcellular Location

Cell junction, Cytoplasm, Nucleus.

UNIPROT

SwissProt: O15117 Human

SwissProt: O35601 Mouse

SwissProt: D3ZIE4 Rat

Synonyms

ADAP antibody

Adhesion and degranulation promoting adaptor protein antibody

FYB 120/130 antibody

Fyb antibody

FYB-120/130 antibody

FYB_HUMAN antibody

FYN binding protein antibody

FYN T binding protein antibody

FYN-binding protein antibody

FYN-T-binding protein antibody

Expand

Images

  • Western blot analysis of FYB on different lysates with Rabbit anti-FYB antibody (<a href="/products/ET7109-38" style="font-weight: bold;text-decoration: underline;">ET7109-38</a>) at 1/5,000 dilution.<br /><br />Lane 1: U-937 cell lysate (15 µg/Lane)<br />Lane 2: Jurkat cell lysate (15 µg/Lane)<br />Lane 3: RAW264.7 cell lysate (15 µg/Lane)<br />Lane 4: Mouse spleen tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 85 kDa<br />Observed band size: 110 kDa<br /><br />Exposure time: 2 minutes; 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/ET7109-38" style="font-weight: bold;text-decoration: underline;">ET7109-38</a>) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 FYB on different lysates with Rabbit anti-FYB antibody (ET7109-38) at 1/5,000 dilution.

    Lane 1: U-937 cell lysate (15 µg/Lane)
    Lane 2: Jurkat cell lysate (15 µg/Lane)
    Lane 3: RAW264.7 cell lysate (15 µg/Lane)
    Lane 4: Mouse spleen tissue lysate (30 µg/Lane)

    Predicted band size: 85 kDa
    Observed band size: 110 kDa

    Exposure time: 2 minutes; 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 (ET7109-38) 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.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-FYB 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 antibody (<a href="/products/ET7109-38" style="font-weight: bold;text-decoration: underline;">ET7109-38</a>) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-FYB 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 antibody (ET7109-38) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human appendix tissue using anti-FYB 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 antibody (<a href="/products/ET7109-38" style="font-weight: bold;text-decoration: underline;">ET7109-38</a>) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human appendix tissue using anti-FYB 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 antibody (ET7109-38) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-FYB 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 antibody (<a href="/products/ET7109-38" style="font-weight: bold;text-decoration: underline;">ET7109-38</a>) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-FYB 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 antibody (ET7109-38) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-FYB antibody (<a href="/products/ET7109-38" style="font-weight: bold;text-decoration: underline;">ET7109-38</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7109-38" style="font-weight: bold;text-decoration: underline;">ET7109-38</a>) at 1/1,000 dilution for 1 hour 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 rat spleen tissue with Rabbit anti-FYB antibody (ET7109-38) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-38) at 1/1,000 dilution for 1 hour 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.

  • Immunocytochemistry analysis of Jurkat cells labeling FYB with Rabbit anti-FYB antibody (<a href="/products/ET7109-38" style="font-weight: bold;text-decoration: underline;">ET7109-38</a>) at 1/100 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-FYB antibody (<a href="/products/ET7109-38" style="font-weight: bold;text-decoration: underline;">ET7109-38</a>) at 1/100 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 Jurkat cells labeling FYB with Rabbit anti-FYB antibody (ET7109-38) at 1/100 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-FYB antibody (ET7109-38) at 1/100 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.

  • Flow cytometric analysis of FYB was done on HepG2 cells. The cells were fixed, permeabilized and stained with FYB antibody at 1/100 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). After incubation of the primary antibody on room temperature for 1 hour, the cells was stained with a Alexa Fluor&trade; 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.

    Flow cytometric analysis of FYB was done on HepG2 cells. The cells were fixed, permeabilized and stained with FYB antibody at 1/100 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). After incubation of the primary antibody on room temperature for 1 hour, the cells was stained with a Alexa Fluor™ 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.

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