Two-color flow cytometric analysis of TIM-4 expression on mouse resident peritoneal exudate cells. C57BL/6 mouse resident peritoneal exudate cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PerCP-Cy™5.5 Rat Anti-Mouse CD11b (Cat. No. 550993/561114) and either Alexa Fluor® 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; Left Panel) or Alexa Fluor® 647 Rat Anti-Mouse TIM-4 antibody (Cat. No. 564178; Right Panel). Two-color flow cytometric contour plots showing the correlated expression patterns of CD11b versus TIM-4 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable resident peritoneal exudate cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of TIM-4 expression on mouse resident peritoneal exudate cells. C57BL/6 mouse resident peritoneal exudate cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PerCP-Cy™5.5 Rat Anti-Mouse CD11b (Cat. No. 550993/561114) and either Alexa Fluor® 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; Left Panel) or Alexa Fluor® 647 Rat Anti-Mouse TIM-4 antibody (Cat. No. 564178; Right Panel). Two-color flow cytometric contour plots showing the correlated expression patterns of CD11b versus TIM-4 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable resident peritoneal exudate cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
BD Pharmingen™
TIMD-4, SMUCKLER, Tim4, Timd4
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2a, κ
Mouse Tim-4 Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
276891
AB_2738647
Aqueous buffered solution containing ≤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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Alexa Fluor® 647 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557690
PerCP-Cy™5.5 Rat Anti-CD11b RUO
Size
25 µg
Cat No.
561114
564178 Rev. 2
Antibody Details
RMT4-54
The RMT4-54 monoclonal antibody specifically binds to TIM-4. TIM-4 is encoded by Timd4 (T cell immunoglobulin and mucin domain containing 4). TIM-4 is also known as Spleen, mucin-containing, knockout of lymphotoxin protein (SMUCKLER). TIM-4 is a single-pass type I membrane transmembrane glycoprotein belonging to the TIM family of the immunoglobulin superfamily. TIM-4 is expressed by macrophages and at low levels by dendritic cells. TIM-4 is a phosphatidylserine receptor that enhances the phagocytosis of apoptotic cells. It can serve as a receptor for TIM-1, also known as the Hepatitis A virus cellular receptor 1 (Havcr1).
564178 Rev. 2
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
564178 Rev.2
Citations & References
Development References (3)
-
Curtiss ML, Gorman JV, Businga TR, et al. Tim-1 regulates Th2 responses in an airway hypersensitivity model. Eur J Immunol. 2012; 42(3):651-661. (Clone-specific: Flow cytometry). 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
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Nakayama M, Akiba H, Takeda K, et al. Tim-3 mediates phagocytosis of apoptotic cells and cross-presentation. Blood. 2009; 113(16):3821-3830. (Immunogen: Blocking, Flow cytometry, Functional assay, Inhibition, In vivo exacerbation). View Reference
564178 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.
