WB
Western blot analysis of VCAM1 on different lysates with Rabbit anti-VCAM1 antibody 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), Exposure time: 24 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-VCAM1 antibody 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 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.IHC
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-VCAM1 antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) 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.ICC/IF
Immunocytochemistry analysis of C2C12 cells labeling VCAM1 with Rabbit anti-VCAM1 antibody 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 at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of C2C12 cells labeling VCAM1. Cells were fixed and permeabilized. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
VCAM1 was immunoprecipitated in 0.2mg C2C12 cell lysate with Rabbit anti-VCAM1 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-VCAM1 antibody 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 Rabbit anti-VCAM1 antibody in C2C12 cell lysate, Lane 3: Rabbit anti-VCAM1 antibody IP in C2C12 cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 5 seconds.| Product Name | VCAM1 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Cell adhesion molecules are a family of closely related cell surface glycoproteins involved in cell-cell interactions during growth and are thought to play an important role in embryogenesis and development. Neuronal cell adhesion molecule (NCAM) expression is observed in a variety of human tumors including neuroblastomas, rhabdomyosarcomas, Wilms' tumors, Ewing's sarcomas and some primitive myeloid malignancies. The intracellular adhesion molecule-1 (ICAM-1), also referred to as CD54, is an integral membrane protein of the immunoglobulin superfamily and recognizes the B2а1 and B2аM integrins. PECAM-1 (platelet/endothelial cell adhesion molecule-1), also referred to as CD31, is a glycoprotein expressed on the cell surfaces of monocytes, neutrophils, platelets and a subpopulation of T cells. VCAM-1 (vascular cell adhesion molecule-1) was first identified as an adhesion molecule induced on human endothelial cells by inflammatory cytokines such as IL-1, tumor necrosis factor (TNF) and lipopolysaccharide (LPS). The KALIG gene encodes a nerve cell adhesion molecule (NCAM) -like protein and is deleted in 66% of patients with Kallmann's syndrome, anosmia with secondary hypogonadism. |
| Immunogen | Synthetic peptide within Human VCAM1 aa 690-739 / 739. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:5000-1:10000 IHC:1:200 ICC/IF:1:1000 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | 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 VCAM-1 antibody VCAM1 antibody VCAM1_HUMAN antibody |
|---|---|
| Molecular Weight(MW) | 81 kDa(Observed band size: 100 kDa) |
| Cellular Localization | Membrane |
WB
Western blot analysis of VCAM1 on different lysates with Rabbit anti-VCAM1 antibody 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), Exposure time: 24 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-VCAM1 antibody 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 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.
IHC
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-VCAM1 antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) 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.
ICC/IF
Immunocytochemistry analysis of C2C12 cells labeling VCAM1 with Rabbit anti-VCAM1 antibody 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 at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of C2C12 cells labeling VCAM1. Cells were fixed and permeabilized. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
VCAM1 was immunoprecipitated in 0.2mg C2C12 cell lysate with Rabbit anti-VCAM1 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-VCAM1 antibody 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 Rabbit anti-VCAM1 antibody in C2C12 cell lysate, Lane 3: Rabbit anti-VCAM1 antibody IP in C2C12 cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 5 seconds.| Application Notes | WB:1:5000-1:10000 IHC:1:200 ICC/IF:1:1000 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
