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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

Validation Data Gallery

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

  • 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.

    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.

Product Information

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

ApplicationsFC, IF
ReactivityHuman
ConjugateCoraLite® Plus 647 Fluorescent Dye
Type Nanobody
ClassRecombinant
HostAlpaca
AffinityPicomolar range, below the assay limit (biolayer interferometry)
Molecular Weight14.824 kDa
FormLiquid
Excitation/Emission Maxima Wavelengths654 nm / 674 nm
RRIDAB_3101922
Storage Buffer500 mM NaCl, 10 mM HEPES pH 7.0, 5 mM EDTA, 0.09% sodium azide
Storage ConditionStore 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).

Documentation

SDS
SDS-Immuno-Oncology VHHs