ChromoTek CoraLite® Plus 647-conjugated TIM3 VHH

CL647-tmt targets TIM3 in FC and IF applications and shows reactivity with Human samples.

Host/Type

Alpaca VHH

Reactivity

Human

Applications

FC, IF

Conjugate

CoraLite® Plus 647 Fluorescent Dye

Cat no : CL647-tmt

Synonyms

TIM3-Nanobody, TIM3-single domain antibody (sdAb), CD366, HAVcr 2, HAVCR2, KIM 3, T cell membrane protein 3, Tim 3, TIM3, TIM-3, TIMD 3, TIMD3

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Validation Data Gallery

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  1X10^6 CHO TIM3 stable expressing cells (red) and CHO parental cells were surface stained with 0.5 µg CoraLite® Plus 647 conjugated-TIM3 VHH (CL647-tmt). Cells were not fixed.

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  1X10^6 CHO TIM3 stable expressing cells (red) were surface stained with 0.5 µg CoraLite® Plus 647 conjugated-TIM3 VHH (CL647-tmt). Cells were not fixed and pre-gated on co-expressed, GFP positive cells.

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  Live HeLa cells transfected with human TIM-3 were immunostained with CoraLite® Plus 555 conjugated-TIM3 VHH (CL555-tmt,1:1000, yellow). After formaldehyde fixation, triton permeabilization and blocking with BSA, intracellular pool of TIM-3 was immunostained with CoraLite® Plus 647 conjugated-TIM3 VHH (CL647-tmt, 1:1000, magenta). Cell nuclei were stained with DAPI (cyan). Epifluorescence images were acquired with a 20x objective and post-processed.

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  Live HeLa cells transfected with human TIM-3 were immunostained with CoraLite® Plus 647 conjugated-TIM3 VHH (CL647-tmt, 1:1000, magenta). Cells were fixed and nuclei were stained with DAPI (cyan). Epifluorescence images were acquired with 20x objective and post-processed.  Note: Immunostaining with CL647-tmt can also be performed after formaldehyde fixation of the cells.

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  Live HeLa cells transfected with human TIM-3 were probed with cobolimab biosimilar (anti-TIM3, huIgG4). After formaldehyde fixation, the cells were immunostained with CoraLite® Plus 647 conjugated-TIM3 VHH (CL647-tmt, 1:1000, magenta) and with anti-human IgG Nano-Secondary (shuGCL488-2, green). Cell nuclei were stained with DAPI (cyan). Epifluorescence images were acquired with 20x objective and post-processed.

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  Left: 1x10^6 PHA-treated (3d) human PBMCs were surface stained with 0.25ug CoraLite® Plus 647 conjugated TIM-3 VHH (CL647-tmt) and CL405 CD3 (CL405-65151). Right: 1x10^6 PHA-treated (3d) human PBMCs were surface stained with 0.25ug CoraLite® Plus 647 conjugated TIM-3 VHH (CL647-tmt) (red) or unstained (blue). 1X10^6 untreated human PBMCs were surface stained with 0.25ug CoraLite® Plus 647 conjugated TIM-3 VHH (CL647-tmt) (black dashed). Cells were treated with FC Receptor Block prior to staining. Cells were not fixed. Lymphocytes were gated.

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Product Information

CL647-tmt targets TIM3 in FC and IF applications and shows reactivity with Human samples.

• Applications: FC, IF
• Reactivity: Human
• Conjugate: CoraLite® Plus 647 Fluorescent Dye
• Type: Nanobody
• Class: Recombinant
• Host: Alpaca
• Affinity: Picomolar range, below the assay limit (biolayer interferometry)
• Molecular Weight: 14.824 kDa
• Form: Liquid
• Excitation/Emission Maxima Wavelengths: 654 nm / 674 nm
• RRID: AB_3101922
• Storage Buffer: 500 mM NaCl, 10 mM HEPES pH 7.0, 5 mM EDTA, 0.09% sodium azide
• Storage Condition: Store at -20°C; Shipped at ambient temperature.
• Background: TIM3, also known as HAVCR2, is a member of the recently discovered T cell Ig and mucin domain-containing molecule superfamily. TIM3 is a negative regulatory molecule that is important for T cell tolerance and has a crucial role in autoimmunity and T cell exhaustion during chronic viral infection (PMID: 23180819). TIM3 is expressed by T-helper type 1 (Th1) cells, macrophage, monocyte, dendritic cells, CD8+ T cell and other lymphocyte subsets. Galectin-9 is a ligand for TIM3. TIM3-galectin-9 pathway negatively regulates T helper type 1 immunity (PMID: 16286920). TIM3 is a 280-aa membrane protein with a calculated molecular weight of 33 kDa, the higher molecular weights between 50 and 70 kDa detected by this monoclonal antibody probably represent glycosylated TIM3 (PMID: 20107545; 17069754; 11725301).