Flow cytometric analysis of Ig κ Light Chain expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were incubated in complete tissue culture medium overnight in order to minimize subsequent nonspecific immunofluorescent staining. The cells were harvested and stained with Alexa Fluor® 488 Mouse anti-Human CD19 (Cat. No. 557697) and either APC-H7 Mouse anti-Human Ig κ Light Chain antibody (Cat. No. 561325; Right Panel) or APC-H7 Mouse IgG1, κ Isotype Control (Cat. No. 560167; Left Panel). The two color flow cytometric dot plots showing the correlated expression of Ig κ Light Chain (or Ig isotype control staining) versus CD19 were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of Ig κ Light Chain expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were incubated in complete tissue culture medium overnight in order to minimize subsequent nonspecific immunofluorescent staining. The cells were harvested and stained with Alexa Fluor® 488 Mouse anti-Human CD19 (Cat. No. 557697) and either APC-H7 Mouse anti-Human Ig κ Light Chain antibody (Cat. No. 561325; Right Panel) or APC-H7 Mouse IgG1, κ Isotype Control (Cat. No. 560167; Left Panel). The two color flow cytometric dot plots showing the correlated expression of Ig κ Light Chain (or Ig isotype control staining) versus CD19 were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
Ig, kappa Light Chain; IGKC; HCAK1; Km; Ig kappa chain C region
Human (QC Testing)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl
3514
AB_10611725
Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
RUO
Preparation And Storage
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 with APC-H7 under optimum conditions, and unconjugated antibody and APC-H7 were removed.
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- BD APC-H7 is a tandem conjugate and an analog of APC-Cy7 with the same spectral properties. It has decreased intensity but it is engineered for greater stability and less spillover in the APC channel and consequently offers better performance than APC-Cy7. It has an absorption maximum of approximately 650 nm. When excited by light from a red laser, the APC fluorochrome can transfer energy to the cyanine dye, which then emits at a longer wavelength. The resulting fluorescent emission maximum is approximately 767 nm. BD recommends that a 750-nm longpass filter be used along with a red-sensitive detector such as the Hamamatsu R3896 PMT. As with APC-Cy7 special filters are required when using APC-H7 in conjunction with APC. Note: Although our APC-H7 products demonstrate higher lot-to lot consistency than other APC tandem conjugate products, and every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-H7 conjugate.
- Although BD APC-H7 is engineered to minimize spillover to the APC channel and is more stable and less affected by light, temperature, and formaldehyde-based fixatives, compared to other APC-cyanine tandem dyes, it is still good practice to minimize as much as possible, any light, temperature and fixative exposure when working with all fluorescent conjugates.
- Cy is a trademark of GE Healthcare.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
APC-H7 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
560167
Alexa Fluor® 488 Mouse Anti-Human CD19 RUO
Size
100 Tests
Cat No.
557697
561325 Rev. 2
Antibody Details
G20-193
The G20-193 monoclonal antibody specifically binds to human immunoglobulin light chain, kappa (κ). It does not react with human immunoglobulin λ light chains or human immunoglobulin heavy chains.
561325 Rev. 2
Format Details
APC-H7
The BD Horizon™ APC-H7 dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of a Allophycocyanin (APC) donor that has excitation maxima (Ex Max) of 659 nm and an acceptor dye, H7, with an emission maximum (Em Max) at 782 nm. APC-H7, driven by BD innovation, is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 780 nm (e.g., a 760/60 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
APC-H7
Red 627-640 nm
659 nm
782 nm
561325 Rev.2
Citations & References
View product citations for antibody "561325" on CiteAb
No Citations are available for this Product
561325 Rev. 2
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.