Product Datasheet
www.leinco.com
excellence in early discovery research       




Goat Anti-Human B-Cell Adhesion Molecule (BCAM)
Antigen Affinity Purified
(BCAM, AU, CD239, LU, MSK19)
Polyclonal Antibody
Share |


Prod. No.:B559
Host:Goat
Pkg. Size:100 µg, 1 mg
Storage:-20°C to -70°C Detailed storage instructions below.
Description
Specificity:
Goat Anti-Human B-Cell Adhesion Molecule (BCAM) recognizes Human BCAM. This antigen affinity purified polyclonal antibody was purified using a proprietary chromatographic technique that includes covalently immobilizing the antigen proteins or peptides to agarose based beads. This purification method enhances specificity, reduces nonspecific binding of extraneous IgG and provides you with the most reliable reagent available for your early discovery research.

Background:
Basal-Cell Adhesion Molecule (BCAM) and Lutheran blood group glycoprotein (LU) are two alternatively spliced variants of a single immunoglobulin superfamily (IgSF) protein that differ in the length of their cytoplasmic tails. BCAM cDNA encodes a 628 amino acid (aa) residues precursor protein with a putative 31 aa signal peptide, a 597 aa extracellular domain containing three C2 type and two V-type Ig-like domains, a 21 aa transmembrane domain, and a 19 aa cytoplasmic domain. Compared to the 40 aa cytoplasmic domain present in LU, the BCAM cytoplasmic tail lacks the putative Src homology 3 (SH3) binding site that may be involved in mediating intracellular signaling. BCAM/LU has wide tissue distribution and is expressed on erythrocytes, the endothelium of blood vessels and on the basal layer of cells in the epithelia. The expression of BCAM/LU in normal tissues is higher in fetal versus adult tissues. BCAM/LU expression is also upregulated in sickle cell disease red blood cells, in activated keratinocytes and following malignant transformation in some cell types in vivo and in vitro. BCAM/LU has been shown to be an adhesion molecule that binds laminin, a basement membrane protein involved in cell differentiation, adhesion, migration and proliferation.

This protein may play a role in epithelial cell cancer and in vaso-occlusion of red blood cells in sickle cell disease. Two transcript variants encoding different isoforms have been found for this gene.

Known Species Cross Reactivity: Hu+
Immunogen
Purified Recombinant Human BCAM (>98%)
Formulation
This antigen affinity purified polyclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco’s phosphate buffered saline (1X PBS) pH 7.2 – 7.3 containing 5.0% w/v trehalose with no calcium, magnesium, or preservatives present.
Reconstitution
We suggest aseptically reconstituting this lyophilized, antigen affinity purified polyclonal antibody with 1.0 µl of sterile 1X PBS pH 7.2-7.3. This will result in a final antibody concentration of 0.1 mg/ml.
Endotoxin
<0.1 EU/µg as determined by the LAL method
Storage and Stability
The lyophilized antigen affinity purified polyclonal antibody can be stored desiccated at -20°C to -70°C for twelve months from date of receipt. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles. No detectable loss of activity was observed after six months.
Reported Applications
Indirect ELISA, WB, B
Indirect ELISA: This antibody can be used in an indirect detection ELISA at 0.5-1.0 µg/ml with a suitable second step reagent such as Donkey Anti-Goat IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: G505). The detection sensitivity of this indirect ELISA for Human BCAM is approximately 0.03 ng/well.
Western Blotting: To detect Human BCAM this polyclonal antibody can be used at a concentration of 0.1-0.2 µg/ml. This polyclonal antibody should be used in conjunction with compatible second-step reagents such as Donkey Anti-Goat IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: G505) and a chromogenic substrate such as TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343). The detection limit for Human BCAM is 0.5 ng/lane under either reducing or non-reducing conditions. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used. A suitable Western blotting control is Recombinant Human BCAM (Leinco Prod. No.: B534).
Blocking: This antibody, at concentrations ≥ 25 µg/ml final, will block the adhesion of human osteosarcoma (ATCC, CRL-1543) cells (0.5 x 106 cells/ml, 100 µl/well) to immobilized human BCAM/Fc (10 µg/ml, 100 µl/well, 1 hour at room temperature).
Each investigator should determine their own optimal working dilution for specific applications.


Country of Origin
USA
Related Products
Donkey Anti-Goat IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: G505)
TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343)
Recombinant Human BCAM (Leinco Prod. No.: B534)
Anti-Human BCAM-Biotin (Antigen Aff. Pur.) (Leinco Prod. No.: B604)
800.538.1145 ® Copyright Leinco Technologies, Inc. 2006 – 2013