Preparation And Storage
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The 1D12 monoclonal antibody specifically binds to CD365, the T-cell immunoglobulin mucin receptor 1 (TIM-1). TIM-1 is expressed on kidney epithelial cells, T cells, and some hematopoietic and non-hematopoietic cells. CD365 (TIM-1) is a type 1 transmembrane glycoprotein that serves as a receptor for hepatitis A virus and is encoded by the HAVCR1 (Hepatitis A virus cellular receptor 1) gene. TIM-1 also serves as a receptor for phosphatidylserine which is exposed on the surface of apoptotic cells. TIM-1 can reportedly mediate the uptake of apoptotic cells through the recognition of phosphatidylserine and thus help maintain tissue homeostasis and self-tolerance. TIM-1 is likewise known as Kidney injury molecule 1 (KIM-1). It is highly expressed by cancerous kidneys, and upregulated in the proximal tubular epithelium and shed into the urine during acute and chronic kidney injury. CD365 (TIM-1) also functions as a costimulatory molecule for immune cells. It is expressed by activated CD4+ T cells and regulates the effector functions (eg, enhanced cytokine production) and survival of differentiated T cells, including those mediating Th2-like immune responses. Other ligands have been described for TIM-1 including TIM-4 and LMIR5 (also known as CD300b) which are expressed by myeloid cells. With respect to disease associations, the HAVCR1 gene has been linked to asthma, allergy, and some autoimmune diseases.
The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.
Development References (7)
-
Binne LL, Scott ML, Rennert PD. Human TIM-1 associates with the TCR complex and up-regulates T cell activation signals. J Immunol. 2007; 178(7):4342-4350. (Biology). View Reference
-
Freeman GJ, Casasnovas JM, Umetsu DT, DeKruyff RH. TIM genes: a family of cell surface phosphatidylserine receptors that regulate innate and adaptive immunity.. Immunol Rev. 2010; 235(1):172-89. (Biology). View Reference
-
Kim HY, Eyheramonho MB, Pichavant M, et al. A polymorphism in TIM1 is associated with susceptibility to severe hepatitis A virus infection in humans. J Clin Invest. 2011; 121(3):1111-1118. (Clone-specific: Blocking, Functional assay). View Reference
-
Kobayashi N, Karisola P, Pena-Cruz V, et al. TIM-1 and TIM-4 glycoproteins bind phosphatidylserine and mediate uptake of apoptotic cells. Immunity. 2007; 27(6):927-940. (Immunogen: Blocking, Flow cytometry, Functional assay). View Reference
-
Manangeeswaran M, Jacques J, Tami C, et al. Binding of hepatitis A virus to its cellular receptor 1 inhibits T-regulatory cell functions in humans. Gastroenterology. 2012; 142(7):1516-1525. (Clone-specific: Blocking, Functional assay). View Reference
-
Rennert PD. Novel roles for TIM-1 in immunity and infection. Immunol Lett. 2011; 141(1):28-35. (Clone-specific: Blocking). View Reference
-
Rodriguez-Manzanet R, DeKruyff R, Kuchroo VK, Umetsu DT. The costimulatory role of TIM molecules. Immunol Rev. 2009; 229(1):259-270. (Biology). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
