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VCAM1 Recombinant Rabbit Monoclonal Antibody [SA05-04]

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Specification

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Catalog# ET1601-18

VCAM1 Recombinant Rabbit Monoclonal Antibody [SA05-04]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

  • FC(Intra)

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Predicted reactivity

  • Zebrafish

Conjugation

  • unconjugated

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

Overview

Product Name

VCAM1 Recombinant Rabbit Monoclonal Antibody [SA05-04]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human VCAM1 aa 690-739 / 739.

Species Reactivity

Human, Mouse, Rat (Predicted: Zebrafish)

Predicted species support after-sales service

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, IP, FC(Intra)

Target Molecular Weight

Predicted band size: 81 kDa

Positive Control

C2C12 cell lysate, 3T3-L1 cell lysate, Mouse spleen tissue lysate, Rat spleen tissue lysate, NIH/3T3 cell lysate, human spleen tissue, C2C12 cell lysate, C2C12.

Conjugation

unconjugated

Clone Number

SA05-04

RRID

AB_3069600

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

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

  • 1:5,000-1:10,000

  • IF-Cell

  • 1:1,000

  • IF-Tissue

  • 1:100

  • IHC-P

  • 1:1,000-1:2,000

  • IP

  • 1-2μg/sample

  • FC(Intra)

  • 1:1,000

Target

Function

Vascular cell adhesion protein 1 also known as vascular cell adhesion molecule 1 (VCAM-1) or cluster of differentiation 106 (CD106) is a protein that in humans is encoded by the VCAM1 gene. VCAM-1 functions as a cell adhesion molecule. The VCAM-1 protein mediates the adhesion of lymphocytes, monocytes, eosinophils, and basophils to vascular endothelium. It also functions in leukocyte-endothelial cell signal transduction, and it may play a role in the development of atherosclerosis and rheumatoid arthritis.

Background References

1. Gregory EK et al. Periadventitial atRA citrate-based polyester membranes reduce neointimal hyperplasia and restenosis after carotid injury in rats. Am J Physiol Heart Circ Physiol 307:H1419-29 (2014).

2. Florea V et al. c-Myc Is Essential to Prevent Endothelial Pro-Inflammatory Senescent Phenotype. PLoS One 8:e73146 (2013).

Tissue Specificity

Expressed on inflamed vascular endothelium, as well as on macrophage-like and dendritic cell types in both normal and inflamed tissue.

Post-translational Modification

Sialoglycoprotein.

Subcellular Location

Membrane

UNIPROT

SwissProt: P19320 Human

SwissProt: P29533 Mouse

SwissProt: P29534 Rat

Synonyms

CD106 antibody

CD106 Antigen antibody

INCAM 100 antibody

INCAM-100 antibody

L1CAM antibody

MGC99561 antibody

V-CAM 1 antibody

Vascular Cell Adhesion Molecule 1 antibody

Vascular cell adhesion protein 1 antibody

VCAM 1 antibody

Expand

Images

  • Western blot analysis of VCAM1 on different lysates with Rabbit anti-VCAM1 antibody (<a href="/products/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: C2C12 cell lysate (20 µg/Lane)<br />Lane 2: 3T3-L1 cell lysate (20 µg/Lane)<br />Lane 3: Mouse spleen tissue lysate (20 µg/Lane)<br />Lane 4: Rat spleen tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 81 kDa<br />Observed band size: 100 kDa<br /><br />Exposure time: 3 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/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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 VCAM1 on different lysates with Rabbit anti-VCAM1 antibody (ET1601-18) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: C2C12 cell lysate (20 µg/Lane)
    Lane 2: 3T3-L1 cell lysate (20 µg/Lane)
    Lane 3: Mouse spleen tissue lysate (20 µg/Lane)
    Lane 4: Rat spleen tissue lysate (20 µg/Lane)

    Predicted band size: 81 kDa
    Observed band size: 100 kDa

    Exposure time: 3 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 (ET1601-18) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of VCAM1 on different lysates with Rabbit anti-VCAM1 antibody (<a href="/products/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a>) at 1/5,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 2: C2C12 cell lysate (20 µg/Lane)<br />Lane 3: RAW264.7 cell lysate (negative) (20 µg/Lane)<br />Lane 4: Mouse spleen tissue lysate (20 µg/Lane)<br />Lane 5: Rat spleen tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 81 kDa<br />Observed band size: 100 kDa<br /><br />Exposure time: 24 seconds; 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/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a>) at 1/5,000 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.

    ☑ Relative expression (RE)

    Western blot analysis of VCAM1 on different lysates with Rabbit anti-VCAM1 antibody (ET1601-18) at 1/5,000 dilution.

    Lane 1: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 2: C2C12 cell lysate (20 µg/Lane)
    Lane 3: RAW264.7 cell lysate (negative) (20 µg/Lane)
    Lane 4: Mouse spleen tissue lysate (20 µg/Lane)
    Lane 5: Rat spleen tissue lysate (20 µg/Lane)

    Predicted band size: 81 kDa
    Observed band size: 100 kDa

    Exposure time: 24 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 (ET1601-18) at 1/5,000 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.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Spleen<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a>, 1/1,000, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Spleen
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: ET1601-18, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Mouse<br />Tissue: Spleen<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a>, 1/1,000, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Spleen
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: ET1601-18, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Rat<br />Tissue: Spleen<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a>, 1/1,000, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Rat
    Tissue: Spleen
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: ET1601-18, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • VCAM1 was immunoprecipitated in 0.2mg C2C12 cell lysate with <a href="/products/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a> at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: C2C12 cell lysate (input)<br />Lane 2: Rabbit IgG instead of <a href="/products/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a> in C2C12 cell lysate<br />Lane 3: <a href="/products/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a> IP in C2C12 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 5 seconds; ECL: <a href="/products/K1802" style="font-weight: bold;text-decoration: underline;">K1802</a>

    VCAM1 was immunoprecipitated in 0.2mg C2C12 cell lysate with ET1601-18 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1601-18 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: C2C12 cell lysate (input)
    Lane 2: Rabbit IgG instead of ET1601-18 in C2C12 cell lysate
    Lane 3: ET1601-18 IP in C2C12 cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 5 seconds; ECL: K1802

  • Immunocytochemistry analysis of C2C12 cells labeling VCAM1 with Rabbit anti-VCAM1 antibody (<a href="/products/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a>) at 1/1,000 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS 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-VCAM1 antibody (<a href="/products/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a>) at 1/1,000 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/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</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 C2C12 cells labeling VCAM1 with Rabbit anti-VCAM1 antibody (ET1601-18) at 1/1,000 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS 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-VCAM1 antibody (ET1601-18) at 1/1,000 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 (M1305-2, 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 C2C12 cells labeling VCAM1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1601-18" style="font-weight: bold;text-decoration: underline;">ET1601-18</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of C2C12 cells labeling VCAM1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-18, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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Citation

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