Description
The 1-155-2 monoclonal antibody specifically recognizes human immunoglobulin lambda light chains (Igλ). Immunoglobulins are expressed on the surface of B lymphocytes and plasma cells, and they also circulate in the serum. The two light chains in an immunoglobulin are of the same type, either kappa (Igκ) or Igλ. Immunoglobulins bearing Igλ are expressed on a minority of normal B lymphocytes and on neoplastic cells of some leukemias, lymphomas, and plasmacytomas. In serum, the 1-155-2 antibody binds to immunoglobulins bearing Igλ as well as free Igλ. It does not bind to Igκ or immunoglobulin heavy chains.
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.
1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
3. For U.S. patents that may apply, see bd.com/patents.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
6. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
7. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
9. An isotype control should be used at the same concentration as the antibody of interest.
10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
11. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
12. Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
13. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Specifications

General

SourceHuman ICAM-3, amino acids Gln30-His485 (Accession # BC058903) with a C-terminal human IgG1 Fc-6His tag was expressed in CHO cells.
Molecular MassThe 701 amino acid recombinant protein has a predicted molecular mass of approximately 77.2 kD. The DTT-reduced protein migrates at approximately 110 kD and non-reduced protein migrate at approximately 220 kD by SDS-PAGE. The predicted N-terminal amino acid is Gln.
Purity>95%, as determined by Coomassie stained SDS-PAGE.
Formulation0.22 µm filtered protein solution is in PBS, pH 7.2
Endotoxin LevelLess than 1.0 EU per µg protein as determined by the LAL method.
Concentration10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
Storage & HandlingUnopened vial can be stored between 2°C and 8°C for up to 2 weeks, at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
ActivityED50= 0.5 - 2.0 µg/mL as measured by the ability of immobilized protein to support the adhesion of retinoic acid (2 µM, 72 hr) activated HL-60 cells.
ApplicationBioassay
Application NotesBioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at [email protected].
StructureHomodimer
DistributionICAM-3 is broadly expressed on leukocytes, endothelial cells, and Langerhans cells
FunctionICAM-3 is ligand for the leukocyte adhesion protein LFA-1 (integrin alpha-L/beta-2) and integrin alpha-D/beta-2. In association with integrin alpha-L/beta-2, it contributes to apoptotic neutrophil phagocytosis by macrophages.
InteractionLeukocytes, endothelial cells
Ligand/ReceptorLFA-1, integrin α-D/β-2
BioactivityMeasured by the ability of immobilized protein to support the adhesion of retinoic acid activated HL-60 cells.
Biology AreaCell Adhesion, Cell Biology, Immunology
Molecular FamilyAdhesion Molecules, CD Molecules, Soluble Receptors
Antigen ReferencesFawcett JH, et al. 1992. Nature. 360:481-484.Vazeux RH, et al. 1992. Nature 360:485-488. Bossy D, et al. 1994. Genomics. 23:712-713. Van der Vieren M, et al. 1995. Immunity. 3:683-690. Starling GC, et al. 1995. Eur J Immunol. 25:2528-2532. Kristof E, et al. 2013. Apoptosis. 18:1235-1251.
Gene ID3385
UniProtView information about ICAM-3 on UniProt.org
Regulatory StatusRUO
Other NamesIntercellular adhesion molecule 3, CD50