WB
Western blot analysis of ICAM1 on different lysates with Rabbit anti-ICAM1 antibody at 1/10,000 dilution. Lane 1: RAW264.7 cell lysate (20 µg/Lane), Lane 2: Mouse spleen tissue lysate (30 µg/Lane), Lane 3: Mouse kidney tissue lysate (30 µg/Lane), Exposure time: 3 minutes; 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 1/10,000 dilution was used in primary antibody dilution 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 mouse lung tissue with Rabbit anti-ICAM1 antibody at 1/2,000 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 at 1/2,000 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.ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling ICAM1 with Rabbit anti-ICAM1 antibody 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-ICAM1 antibody at 1/200 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.IF-F
Species: Mouse, Site: small intestine, Sample: Frozen section, Antibody concentration: 1/1,000, Antigen retrieval: Not required.IF-P
Species: Mouse, Tissue: Lung, Sample: Paraffin-embedded section, Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃. Wash buffer: 1× TBST, Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature. Primary antibody: 1/500, overnight at 4℃. Secondary antibody: Goat Anti-Rabbit IgG (488), 1.5 hours at room temperature.FC
Flow cytometric analysis of mouse spleen cells labeling ICAM1. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (1/2,000) (red). 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
ICAM1 was immunoprecipitated from 0.2 mg mouse spleen tissue lysate with Rabbit anti-ICAM1 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-ICAM1 antibody at 1/10,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Mouse spleen tissue lysate (input), Lane 2: Rabbit anti-ICAM1 antibody IP in mouse spleen tissue lysate, Lane 3: Rabbit IgG instead of Rabbit anti-ICAM1 antibody in mouse spleen tissue lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 59 seconds.| Product Name | ICAM1 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within mouse ICAM1 aa 1-485. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIF-FIF-PIHCIPWB |
| WB:1:1000 IHC:1:2000 ICC/IF:1:200 IF-F:1:1000 IF-P:1:500 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | Mouse |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | ICAM1 |
|---|---|
| Gene Synonyms | BB2 CD54 P3.58 |
| Gene Full Name | intercellular adhesion molecule 1 |
| Gene Summary | This gene encodes a cell surface glycoprotein which is typically expressed on endothelial cells and cells of the immune system. It binds to integrins of type CD11a / CD18, or CD11b / CD18 and is also exploited by Rhinovirus as a receptor. [provided by RefSeq, Jul 2008] |
| Alternative Names | Antigen identified by monoclonal antibody BB2 antibody BB 2 antibody BB2 antibody CD 54 antibody CD_antigen=CD54 antibody CD54 antibody Cell surface glycoprotein P3.58 antibody Human rhinovirus receptor antibody ICAM 1 antibody Antigen identified by monoclonal antibody BB2 antibody BB 2 antibody BB2 antibody CD 54 antibody CD_antigen=CD54 antibody CD54 antibody Cell surface glycoprotein P3.58 antibody Human rhinovirus receptor antibody ICAM 1 antibody ICAM-1 antibody ICAM1 antibody ICAM1_HUMAN antibody intercellular adhesion molecule 1 (CD54) human rhinovirus receptor antibody Intercellular adhesion molecule 1 antibody Major group rhinovirus receptor antibody MALA 2 antibody MALA2 antibody MyD 10 antibody MyD10 antibody P3.58 antibody Surface antigen of activated B cells BB2 antibody |
| Molecular Weight(MW) | 60kDa(Observed band size: 120kDa) |
| Cellular Localization | Membrane. |
WB
Western blot analysis of ICAM1 on different lysates with Rabbit anti-ICAM1 antibody at 1/10,000 dilution. Lane 1: RAW264.7 cell lysate (20 µg/Lane), Lane 2: Mouse spleen tissue lysate (30 µg/Lane), Lane 3: Mouse kidney tissue lysate (30 µg/Lane), Exposure time: 3 minutes; 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 1/10,000 dilution was used in primary antibody dilution 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 mouse lung tissue with Rabbit anti-ICAM1 antibody at 1/2,000 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 at 1/2,000 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.
ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling ICAM1 with Rabbit anti-ICAM1 antibody 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-ICAM1 antibody at 1/200 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.
IF-F
Species: Mouse, Site: small intestine, Sample: Frozen section, Antibody concentration: 1/1,000, Antigen retrieval: Not required.
IF-P
Species: Mouse, Tissue: Lung, Sample: Paraffin-embedded section, Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃. Wash buffer: 1× TBST, Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature. Primary antibody: 1/500, overnight at 4℃. Secondary antibody: Goat Anti-Rabbit IgG (488), 1.5 hours at room temperature.
FC
Flow cytometric analysis of mouse spleen cells labeling ICAM1. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (1/2,000) (red). 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
ICAM1 was immunoprecipitated from 0.2 mg mouse spleen tissue lysate with Rabbit anti-ICAM1 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-ICAM1 antibody at 1/10,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Mouse spleen tissue lysate (input), Lane 2: Rabbit anti-ICAM1 antibody IP in mouse spleen tissue lysate, Lane 3: Rabbit IgG instead of Rabbit anti-ICAM1 antibody in mouse spleen tissue lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 59 seconds.| Application Notes | WB:1:1000 IHC:1:2000 ICC/IF:1:200 IF-F:1:1000 IF-P:1:500 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
