VCAM1 Rabbit pAb

Sample: VCAM1 protein (Human) at 100 ng Primary: Anti-VCAM-1 (AP93960) at 1/300 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 81 kD Observed band size: 81 kD
Tissue/cell: human lung carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min; Incubation: Anti-VCAM-1 Polyclonal Antibody, Unconjugated(AP93960) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Blank control: Mouse Spleen(blue). Primary Antibody:Rabbit Anti-VCAM-1 antibody (AP93960,Green); Dilution: 1 µg in 100 µL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions; Secondary Antibody: Goat anti-rabbit IgG-FITC(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA. Protocol The cells were fixed with 2% paraformaldehyde for 10 min at 37℃. Primary antibody (AP93960, 1 µg /8x10^5 cells) were incubated for 30 min at room temperature, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (1 hour) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/FITC antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 40 min at room temperature. Acquisition of 20,000 events was performed.
Sample: Lane 1: Siha (Human) Cell Lysate at 30 ug Lane 2: Huvec (Human) Cell Lysate at 30 ug Lane 3: U251 (Human) Cell Lysate at 30 ug Primary: Anti-VCAM-1 (AP93960) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 110 kD Observed band size: 110 kD
Sample: Lane 1: Mouse Kidney tissue lysates Lane 2: Mouse Spleen tissue lysates Lane 3: Mouse Cerebrum tissue lysates Primary: Anti-VCAM1 (AP93960) at 1/500 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 81 kDa Observed band size: 100 kDa

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC-P, IHC-F, IF, E
Reactivity	Mouse
Host	Rabbit
Clonality	Polyclonal
Calculated MW	81 KDa
Physical State	Liquid
Immunogen	Recombinant Mouse VCAM 1 protein
Isotype	IgG
Purity	affinity purified by Protein A
Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
SUBCELLULAR LOCATION	Isoform 1: Cell membrane; Single-pass type I membrane protein. Isoform 2: Cell membrane; Lipid-anchor, GPI-anchor.
SIMILARITY	Contains 7 Ig-like C2-type (immunoglobulin-like) domains.
SUBUNIT	Binds to ECMV-D capsid proteins and acts as a receptor for this virus.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Background Descriptions	VCAM1 is important in cell-cell recognition. Appears to function in leukocyte-endothelial cell adhesion. Interacts with the integrins alpha4 beta1 (beta 1 integrin VLA4) and alpha4 beta7 on leukocytes, and mediates both adhesion and signal transduction. The VCAM1/VLA4 interaction may play a pathophysiologic role both in immune responses and in leukocyte emigration to sites of inflammation. VCAM1 is also expressed by several non endothelial cell types including some macrophages, follicular dendritic cells and bone marrow, stromal cells.

Additional Information
Target/Specificity	Expressed on inflamed vascular endothelium, as well as on macrophage-like and dendritic cell types in both normal and inflamed tissue. Expressed in the bone marrow.
Dilution	WB=1:500-2000,IHC-P=1:100-500,IHC-F=1:100-500,IF=1:100-500,Flow-Cyt=1 µg/Test,ELISA=1:5000-10000
Format	0.01M TBS(pH7.4) with 1% BSA, 0.09% (W/V) sodium azide and 50% Glyce
Storage	Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.

Research Areas

For Research Use Only. Not For Use In Diagnostic Procedures.

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol