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Brilliant Violet 421™ anti-mouse NK-1.1 Antibody

Pricing & Availability
Clone
PK136 (See other available formats)
Regulatory Status
RUO
Other Names
NKR-P1C, NKR-P1B, Ly-55, CD161, CD161b, CD161c
Isotype
Mouse IgG2a, κ
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Product Citations
publications
1_PK136_BV421_1_032911
C57BL/6 mouse splenocytes were stained with CD49b/DX5 PE and NK1.1 (clone PK136) Brilliant Violet 421™ (top) or mouse IgG2a, κ Brilliant Violet 421™ isotype control (bottom).
  • 1_PK136_BV421_1_032911
    C57BL/6 mouse splenocytes were stained with CD49b/DX5 PE and NK1.1 (clone PK136) Brilliant Violet 421™ (top) or mouse IgG2a, κ Brilliant Violet 421™ isotype control (bottom).
  • 2_PK136_BV421_2_032911
  • 3_53_Mouse_Liver_CD8_CD44_NK1.1
    Confocal image of C57BL/6 mouse liver sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD8 (cyan) in Cycle 2, CD44 (blue) in Cycle 2, and NK1.1 (red) in Cycle 3. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
  • 4_59_Mouse_Lymph_Node_F480_CD68_NK1.1
    Mice were injected subcutaneously with sheep red blood cells in a volume of 25 µl per site on days 0 and 4 and harvested on day 11. Confocal image of C57BL/6 mouse lymph node acquired using the IBEX method of highly multiplexed antibody-based imaging: F4/80 (cyan) in Cycle 3, CD68 (blue) in Cycle 6, and NK1.1 (magenta) in Cycle 9. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
See Brilliant Violet 421™ spectral data
Cat # Size Price Quantity Check Availability Save
108731 125 µL £133
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108741 50 µg £163
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108732 500 µL £271
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Description

NK-1.1 surface antigen, also known as CD161b/CD161c and Ly-55, is encoded by the NKR-P1B/NKR-P1C gene. It is expressed on NK cells and NK-T cells in some mouse strains, including C57BL/6, FVB/N, and NZB, but not AKR, BALB/c, CBA/J, C3H, DBA/1, DBA/2, NOD, SJL, and 129. Expression of NKR-P1C antigen has been correlated with lysis of tumor cells in vitro and rejection of bone marrow allografts in vivo. NK-1.1 has also been shown to play a role in NK cell activation, IFN-γ production, and cytotoxic granule release. NK-1.1 and DX5 are commonly used as mouse NK cell markers.

Product Details
Technical Data Sheet (pdf)

Product Details

Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
NK-1+ cells from mouse spleen and bone marrow
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA).
Preparation
The antibody was purified by affinity chromatography and conjugated with Brilliant Violet 421™ under optimal conditions.
Concentration
µg sizes: 0.2 mg/mL
µL sizes: lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC - Quality tested

SB - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining using the µg size, the suggested use of this reagent is ≤ 0.5 µg per million cells in 100 µl volume. For immunofluorescent staining using µl sizes, the suggested use of this reagent is 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.

Brilliant Violet 421™ excites at 405 nm and emits at 421 nm. The standard bandpass filter 450/50 nm is recommended for detection. Brilliant Violet 421™ is a trademark of Sirigen Group Ltd.


Learn more about Brilliant Violet™.

This product is subject to proprietary rights of Sirigen Inc. and is made and sold under license from Sirigen Inc. The purchase of this product conveys to the buyer a non-transferable right to use the purchased product for research purposes only. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics or diagnostics is strictly prohibited. This product is covered by U.S. Patent(s), pending patent applications and foreign equivalents.
Excitation Laser
Violet Laser (405 nm)
Application Notes

Additional reported applications (for the relevant formats) include: immunoprecipitation1,2, complement-dependent cytotoxicity3, in vivo depletion4,5,9,10, mediation of in vitro redirected lysis6, blocking of NK cell function7, induction of proliferation8, immunohistochemical staining of frozen sections11, immunofluorescence microscopy11, and spatial biology (IBEX)16,17. The LEAF™ purified antibody (Endotoxin <0.1 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 108712).

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

Application References

(PubMed link indicates BioLegend citation)
  1. Carlyle JR, et al. 1999. J. Immunol. 162:5917. (IP)
  2. Sentman CL, et al. 1989. Hybridoma 8:605. (IP)
  3. Koo GC, et al. 1984. Hybridoma 3:301. (Cyt)
  4. Sentman CL, et al. 1989. J. Immunol. 142:1847. (Deplete)
  5. Koo GC, et al. 1986. J. Immunol. 137:3742. (Deplete)
  6. Karlhofer FM, et al. 1991. J. Immunol. 146:3662.
  7. Kung SK, et al. 1999. J. Immunol. 162:5876. (Block)
  8. Reichlin A, et al. 1998. Immunol. Cell Biol. 76:143.
  9. Drobyski W, et al. 1996. Blood 87:5355. (Deplete)
  10. Andoniou CE, et al. 2005. Nat. Immunol. 6:1011. (Deplete)
  11. Kanwar JR, et al. 2001. J. Natl. Cancer Inst. 93:1541. (IHC, IF)
  12. Kroemer A, et al. 2008. J. Immunol. 180:7818. PubMed
  13. Kim JY, et al. 2009. Exp Mol Med. 30:288. PubMed
  14. Bankoti J, et al. 2010. Toxicol. Sci. 115:422. (FC) PubMed
  15. Lee H, et al. 2014. Invest Ophthalmol Vis Sci. 55:2885. PubMed
  16. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  17. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Li Q et al. 2018. Immunity. 48(2):258-270 . PubMed
  2. Baomei Wang et al. 2019. Cell reports. 26(6):1614-1626 . PubMed
  3. Dyer DP et al. 2019. Immunity. 50(2):378-389 . PubMed
  4. Minutti CM, et al. 2019. Immunity. 50:645. PubMed
  5. LaFleur MW, et al. 2019. Nat Commun. 10:1668. PubMed
  6. Schmidleithner L et al. 2019. Immunity. 50(5):1232-1248 . PubMed
  7. Santecchia I, et al. 2019. PLoS Pathog. 15:e1007811. PubMed
  8. Collins PL et al. 2018. Cell. 176(1-2):348-360 . PubMed
  9. Liu H et al. 2017. Cell host & microbe. 22(5):653-666 . PubMed
  10. Alexander Mildner et al. 2017. Immunity. 46(5):849-862 . PubMed
  11. Garber C, et al. 2019. Nat Neurosci. 1.802777778. PubMed
  12. Adams RCM, et al. 2019. Mediators Inflamm. 2019:9160941. PubMed
  13. Bhattacherjee A, et al. 2019. Commun Biol. 2:450. PubMed
  14. Renner K, et al. 2020. Cell Reports. 29(1):135-150.e9.. PubMed
  15. Rasid O, et al. 2020. Cell Reports. 29(12):3933-3945.e3.. PubMed
  16. Gordon E, et al. 2015. Proc Natl Acad Sci U S A. 112: 13075 - 13080. PubMed
  17. Damgaard RB et al. 2016. Cell. 166(5):1215-1230 . PubMed
  18. Lin J, et al. 2017. Sci Rep. 7:41722. PubMed
  19. Bayik D, et al. 2020. Cancer Discov. 1.256944444. PubMed
  20. Wang J, et al. 2020. Cell. 183(7):1867-1883.e26. PubMed
  21. Wiesner DL, et al. 2020. Cell Host Microbe. 614:27. PubMed
  22. Godbersen-Palmer C, et al. 2020. J Immunol. 204:2973. PubMed
  23. Garcia LR, et al. 2021. Nat Commun. 12:3364. PubMed
  24. Tomita T, et al. 2021. Nat Commun. 12:3655. PubMed
  25. Zhao Z, et al. 2021. Nat Commun. 12:4355. PubMed
  26. Machata S, et al. 2021. Front Immunol. 11:565869. PubMed
  27. Wang F, et al. 2021. Cell Mol Gastroenterol Hepatol. 13:257. PubMed
  28. Littwitz-Salomon E, et al. 2021. Nat Commun. 12:5376. PubMed
  29. Chang MH, et al. 2021. Cell Rep. 37:109902. PubMed
  30. Boulch M, et al. 2021. Sci Immunol. 6:. PubMed
  31. Hakim R, et al. 2021. J Neurosci. 41:8441. PubMed
  32. Duan H, et al. 2021. J Clin Invest. 131:. PubMed
  33. Dhayade S, et al. 2020. Nutrients. 12:. PubMed
  34. Hu M, et al. 2020. Cancer Immunol Res. 8:1150. PubMed
  35. Otvos B, et al. 2021. Clin Cancer Res. 27:2038. PubMed
  36. Patil ND, et al. 2022. Front Immunol. 13:818015. PubMed
  37. Saber M, et al. 2021. J Neurosci Res. 99:1136. PubMed
  38. Yang P, et al. 2022. Nat Commun. 13:5782. PubMed
  39. Kim DK, et al. 2022. Nat Commun. 13:6292. PubMed
  40. Jolly A, et al. 2022. Cell Rep Methods. 2:100315. PubMed
  41. Gomez S, et al. 2022. J Immunother Cancer. 10:. PubMed
  42. Schnell A, et al. 2021. Cell. 184:6281. PubMed
  43. Bettke JA, et al. 2022. Infect Immun. 90:e0007022. PubMed
  44. Edwards SC, et al. 2023. J Exp Med. 220: . PubMed
  45. Nettersheim FS, et al. 2023. Front Cardiovasc Med. 9:1076808. PubMed
  46. Tao X, et al. 2022. J Exp Med. 219:. PubMed
  47. Harapas CR, et al. 2022. Sci Immunol. 7:eabi4611. PubMed
  48. Cheng C, et al. 2022. Cell Mol Gastroenterol Hepatol. 15:261. PubMed
  49. Seclì L, et al. 2023. J Immunother Cancer. 11:. PubMed
RRID
AB_10895916 (BioLegend Cat. No. 108731)
AB_2562561 (BioLegend Cat. No. 108741)
AB_2562218 (BioLegend Cat. No. 108732)

Antigen Details

Structure
NKR-P1 gene family
Distribution

NK and NK-T cells in the NK1.1 mouse strains (C57BL, FVB/N, NZB)

Function
NK cell activation, IFN-γ production, cytotoxic granule release
Cell Type
NK cells, NKT cells
Biology Area
Immunology, Innate Immunity
Antigen References

1. Lanier LL. 1997. Immunity 6:371.
2. Yokoyama WM, et al. 1993. Ann. Rev. Immunol. 11:613.
3. Koo GC, et al. 1986. J. Immunol. 137:3742.
4. Giorda R, et al. 1991. J. Immunol. 147:1701.

Gene ID
17059 View all products for this Gene ID
UniProt
View information about NK-1.1 on UniProt.org

Related FAQs

What is the F/P ratio range of our BV421™ format antibody reagents?

It is lot-specific. On average it ranges between 2-4.

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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