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Integrin alpha 5 Recombinant Rabbit Monoclonal Antibody [JJ08-94]

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Specification

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Catalog# ET1701-58

Integrin alpha 5 Recombinant Rabbit Monoclonal Antibody [JJ08-94]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Mouse

  • Green monkey

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

Integrin alpha 5 Recombinant Rabbit Monoclonal Antibody [JJ08-94]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within C-terminal human Integrin alpha 5.

Species Reactivity

Human, Mouse, Green monkey, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, IP, FC

Molecular Weight

Predicted band size: 115 kDa

Positive Control

HeLa cell lysate, A549 cell lysate, U-937 cell lysate, NIH/3T3 cell lysate, COS-1 cell lysate, Mouse liver tissue lysate, Rat liver tissue lysate, NIH/3T3, human kidney tissue, human placenta tissue, human tonsil tissue, mouse endometrium tissue, Jurkat.

Conjugation

unconjugated

Clone Number

JJ08-94

RRID

AB_3068339

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). 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

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:200-1:1,000

  • FC

  • 1:50-1:100

  • IP

  • Use at an assay dependent concentration.

Target

Function

Integrins are heterodimers composed of noncovalently associated transmembrane α and β subunits. The 16 α and 8 β subunits heterodimerize to produce more than 20 different receptors. Most integrin receptors bind ligands that are components of the extracellular matrix, including Fibronectin, Collagen and Vitronectin. Certain integrins can also bind to soluble ligands such as Fibrinogen, or to counterreceptors on adjacent cells such as the intracellular adhesion molecules (ICAMs), leading to aggregation of cells. Ligands serve to cross-link or cluster integrins by binding to adjacent integrin receptors; both receptor clustering and ligand occupancy are necessary for the activation of integrin-mediated responses. In addition to mediating cell adhesion and cytoskeletal organization, integrins function as signaling receptors. Signals transduced by integrins play a role in many biological processes, including cell growth, differentiation, migration and apoptosis.

Background References

1. Shen, Y. et al. 2015. Effect of surface chemistry on the integrin induced pathway in regulating vascular endothelial cells migration. Colloids and surfaces. B, Biointerfaces. 126: 188-97.

2. Elloumi-Hannachi, I. et al. 2015. Contributions of the integrin β1 tail to cell adhesive forces. Experimental cell research. 332: 212-22.

Sequence Similarity

Belongs to the integrin alpha chain family.

Post-translational Modification

Proteolytic cleavage by PCSK5 mediates activation of the precursor.

Subcellular Location

Membrane, Cell junction, Cell surface.

UNIPROT

SwissProt: P08648 Human

SwissProt: P11688 Mouse

Unigene:100796 Rat

Synonyms

CD49 antigen-like family member E antibody

CD49e antibody

Fibronectin receptor subunit alpha antibody

Fibronectin receptor, alpha subunit antibody

FNRA antibody

Integrin alpha 5 (fibronectin receptor alpha) antibody

Integrin alpha-5 antibody

Integrin alpha-5 light chain antibody

Integrin alpha-F antibody

Integrin, alpha 5 (fibronectin receptor, alpha polypeptide) antibody

Expand

Images

  • Western blot analysis of Integrin alpha 5 on different lysates with Rabbit anti-Integrin alpha 5 antibody (<a href="/products/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: A549 cell lysate<br />Lane 3: U-937 cell lysate<br />Lane 4: NIH/3T3 cell lysate<br />Lane 5: COS-1 cell lysate<br />Lane 6: Mouse liver tissue lysate<br />Lane 7: Rat liver tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 115 kDa<br />Observed band size: 140/19 kDa<br /><br />Exposure time: 1 minute 2 seconds; ECL: K1801;<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/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</a>) at 1/1,000 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 Integrin alpha 5 on different lysates with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: A549 cell lysate
    Lane 3: U-937 cell lysate
    Lane 4: NIH/3T3 cell lysate
    Lane 5: COS-1 cell lysate
    Lane 6: Mouse liver tissue lysate
    Lane 7: Rat liver tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 115 kDa
    Observed band size: 140/19 kDa

    Exposure time: 1 minute 2 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 (ET1701-58) at 1/1,000 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;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Integrin alpha 5 on different lysates with Rabbit anti-Integrin alpha 5 antibody (<a href="/products/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Integrin alpha 5 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 115 kDa<br />Observed band size: 19 kDa<br /><br />Exposure time: 2 minutes; ECL: K1801;<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/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</a>) at 1/2,000 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.

    ☑ Knockdown (KD)

    Western blot analysis of Integrin alpha 5 on different lysates with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Integrin alpha 5 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 115 kDa
    Observed band size: 19 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 (ET1701-58) at 1/2,000 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.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling Integrin alpha 5 with Rabbit anti-Integrin alpha 5 antibody (<a href="/products/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</a>) at 1/200 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-Integrin alpha 5 antibody (<a href="/products/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</a>) at 1/200 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 NIH/3T3 cells labeling Integrin alpha 5 with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) at 1/200 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-Integrin alpha 5 antibody (ET1701-58) at 1/200 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.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Integrin alpha 5 antibody (<a href="/products/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</a>) at 1/200 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/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</a>) at 1/200 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 human kidney tissue with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) at 1/200 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 (ET1701-58) at 1/200 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 human placenta tissue with Rabbit anti-Integrin alpha 5 antibody (<a href="/products/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</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/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</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 human placenta tissue with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) 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 (ET1701-58) 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 human tonsil tissue with Rabbit anti-Integrin alpha 5 antibody (<a href="/products/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</a>) at 1/200 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/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</a>) at 1/200 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 human tonsil tissue with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) at 1/200 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 (ET1701-58) at 1/200 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 mouse endometrium tissue with Rabbit anti-Integrin alpha 5 antibody (<a href="/products/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</a>) at 1/200 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/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</a>) at 1/200 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 mouse endometrium tissue with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) at 1/200 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 (ET1701-58) at 1/200 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.

  • Flow cytometric analysis of Integrin alpha 5 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1701-58" style="font-weight: bold;text-decoration: underline;">ET1701-58</a>, 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).

    Flow cytometric analysis of Integrin alpha 5 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1701-58, 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).

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