Description
The 107-1A4 monoclonal antibody specifically recognizes human Prostate-Specific Membrane Antigen (PSMA) which is also known as Glutamate carboxypeptidase II (GCPII), and N-acetyl-L-aspartyl-L-glutamate peptidase I (NAALADase I). PSMA is an ~100 kDa type II transmembrane glycoprotein. It is comprised of a large extracellular region with protease, apical and C-terminal domains that function in substrate binding and enzymatic activity, a transmembrane region, and a short intracellular N-terminal domain. PSMA is encoded by FOLH1 (Folate hydrolase 1) which belongs to the M28 peptidase family. PSMA is expressed in the brain and by cells in various tissues including the prostate, intestines, testis, bladder, liver, kidney, breast, and ovary. In the intestines, this enzyme functions as a folate hydrolase that cleaves glutamate from dietary folates to facilitate the cellular uptake of folic acid. Through its NAALADase activity, this enzyme also acts in the nervous system to hydrolyze the N-aceylaspartylglutamate (NAAG) neuropeptide into NAA and glutamate. This enzyme can thereby modulate excitatory neurotransmission with the release of glutamate which serves as an excitatory neurotransmitter. Overexpression of PSMA by cancer cells, such as prostate cancer cells, may allow for the increased uptake of folates required for rapid cellular division leading to tumor progression. PSMA expression has also been found by some urothelial adenocarcinoma cells and associated with the tumor neovasculature of some tumors as well. The 107-1A4 antibody reportedly recognizes a distinct conformational epitope in the PSMA extracellular domain.
Bead-based compensation or unmixing controls, such as BD® CompBeads or BD™ SpectraComp™, can be used as surrogates to assess fluorescence spillover when bound to fluorochrome-conjugated antibodies. Although these beads have spectral properties similar to cells, variations in spectral emission may occur, resulting in differing spillover values compared to biological controls. Therefore, it is considered best practice to compare the spillover obtained from cells and bead-based controls when using BD® CompBeads or BD™ SpectraComp™ for the first time, to ensure they are appropriate for the intended 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.
