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MABF245

Sigma-Aldrich

Anti-TIM-1 Antibody, clone 3B3

clone 3B3, from rat

Synonym(s):

Hepatitis A virus cellular receptor 1 homolog, HAVcr-1, Kidney injury molecule 1, KIM-1, T cell immunoglobulin and mucin domain-containing protein 1, T-cell immunoglobulin mucin receptor 1, T cell membrane protein 1, TIM-1, TIMD-1

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702

biological source

rat

Quality Level

antibody form

purified antibody

antibody product type

primary antibodies

clone

3B3, monoclonal

species reactivity

mouse

technique(s)

flow cytometry: suitable

isotype

IgG2aκ

NCBI accession no.

UniProt accession no.

shipped in

ambient

target post-translational modification

unmodified

Gene Information

human ... HAVCR1(26762)

General description

Hepatitis A virus cellular receptor 1 homolog (UniProt Q5QNS5; also known as HAVcr-1, Kidney injury molecule 1, KIM-1, T cell immunoglobulin and mucin domain-containing protein 1, T-cell immunoglobulin mucin receptor 1, T cell membrane protein 1, TIM-1, TIMD-1) is encoded by the Havcr1 (also known as Kim1, Tim1, Timd1) gene (Gene ID 171283) in murine species. TIM‐1/KIM‐1 is a phosphatidylserine phagocytosis and scavenger receptor expressed on kidney proximal tubule epithelial cells and, at a much lower level, also among a subset of immune cells. Upon kidney injury, TIM‐1 is upregulated more than a thousand fold and assists tissue injury recovery through phagocytosis of apoptotic cells and cellular debris. KIM‐1‐mediated phagocytosis leads to pro‐tolerogenic antigen presentation, which suppresses CD4+ T‐cell proliferation and increases the percentage of regulatory T-cells in an autophagy gene‐dependent manner. Phagocytosis mediated by KIM‐1 in epithelial cells therefore limits inflammation by both removing dead cells and inducing anti‐inflammatory Treg signaling. Human HAVCR1 polymorphisms are linked to various immunological disorders including allergy, asthma, rheumatoid arthritis, atopic dermatitis, and lupus. TIM-1 is produced with a signal peptide (a.a. 1-21) sequence which is removed posttranslationally to yield the mature protein with a large extracellular region (a.a. 21-290 in human and 22-237 in mouse) composed of a V-type Ig-like domain (a.a. 21-121 in human and 22-122 in mouse) and a mucin domain (the rest of the extracellular region), followed by a transmembrane domain (a.a. 238-258), and a cytoplasmic tail (a.a. 312-359 in human and 259-305 in mouse) with tyrosine phosphorylation sites.

Specificity

Clone 3B3 detected recombinant murine TIM-1, but not TIM-3 or TIM-4, Ig fusion. Clone 3B3 immunostained the surface of TIM-1-transfected, but not untransfected 300.19 mouse pre-B-cells (Umetsu, S.E., et al. (2005). Nat. Immunol.6(5):447-454). Clone 3B3 and clone RMT1-10 (Cat. No. MABF225) exhibited opposite effects toward T cell antigen responses (Xiao, S., et al. (2007). J. Exp. Med. 204(7):1691-1702).

Immunogen

Recombinant TIM-1 Ig fusion consists of mouse TIM-1 signal and IgV domains linked to the hinge-CH2-CH3 domains of mouse IgG2a (Umetsu, S.E., et al. (2005). Nat. Immunol.6(5):447-454).

Application

Affects Function: A representative lot exhibited opposite effects toward T cell antigen responses. Clone 3B3 enhanced, while RMT1-10 decreased antigen-dependent T cell proliferation (Xiao, S., et al. (2007). J. Exp. Med. 204(7):1691-1702).

Affects Function: A representative lot induced a rapid capping of anti-CD3 bound CD3 on the surface of CD4+ T cells with a concomitant T cell morphology change and increased motility (Xiao, S., et al. (2007). J. Exp. Med. 204(7):1691-1702).

Affects Function: A representative lot enhanced the severity of experimental autoimmune encephalomyelitis (EAE) in mice, while clone RMT1-10 prevented EAE (Xiao, S., et al. (2007). J. Exp. Med. 204(7):1691-1702).

Affects Function: A representative lot enhanced T cell proliferation by crosslinking TIM-1 in vitro. When injected in mice in vivo at the beginning and during OVA sensitization, clone 3B3 prevented tolerance induction and enhanced response of isolated spleen T cells to OVA stimulation in culture (Umetsu, S.E., et al. (2005). Nat. Immunol.6(5):447-454).

Affinity Binding Assay: A representative lot bound TIM-1 with a similar association rate as clone RMT1-10 (Cat. No. MABF225), while clone 3B3 displayed a 10-times slower dissociation rate than clone RMT1-10 (Xiao, S., et al. (2007). J. Exp. Med. 204(7):1691-1702).

ELISA Analysis: Representative lots detected recombinant murine TIM-1, but not TIM-3 or TIM-4, Ig fusion protein. Clone 3B3 displayed an enhanced affinity toward TIM-4 fusion construct lacking the mucin domain when compared to full-length TIM-4 fusion construct (Xiao, S., et al. (2007). J. Exp. Med. 204(7):1691-1702; Umetsu, S.E., et al. (2005). Nat. Immunol.6(5):447-454).

Flow Cytometry Analysis: A representative lot immunostained the surface of TIM-1-transfected, but not untransfected or TIM-2-transfected 300.19 mouse pre-B-cells. A time-dependent increase of CD4+ cell surface TIM-1 expression was detected following TCR activation by APCs or by anti-CD3 and anti-CD28 antibodies (Umetsu, S.E., et al. (2005). Nat. Immunol.6(5):447-454).
Research Category
Inflammation & Immunology
This rat monoclonal Anti-TIM-1, clone 3B3 Antibody, Cat. No. MABF245 is validated for use in Affinity Binding Assay, ELISA, Flow Cytometry, and Function analysis of mouse TIM-1.

Quality

Evaluated by Flow Cytometry in murine TIM-1-transfected 300.19 mouse pre-B-cells.

Flow Cytometry Analysis: 0.5 µg of this antibody detected the exogenously expressed murine TIM-1 on the surface of one million transfected 300.19 mouse pre-B-cells.

Target description

31.15/28.72 kDa (mature isoform 1/2) and 33.36/30.93 kDa (isoform 1/2 pro-form) calculated.

Physical form

Format: Purified
Protein G purified.
Purified rat IgG2a in buffer containing PBS without preservatives.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

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