Human/Mouse/Rat TRAM/TICAM2 Antibody

Name: Human/Mouse/Rat TRAM/TICAM2 Antibody
Brand: R&D Systems
SKU: AF4348
Price: 389 USD

Catalog #: AF4348 Datasheet / COA / SDS

Discontinued Product

AF4348 has been discontinued.

View all TRAM/TICAM2 products.

Detection of Human/Mouse/Rat TRAM/TICAM2 by Western Blot.

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Human/Mouse/Rat TRAM/TICAM2 Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects endogenous human, mouse and rat TRAM/TICAM2 in Western blots.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant mouse TRAM/TICAM2
Met1-Ala232
Accession # Q8BJQ4

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

5-15 µg/mL

See below

Intracellular Staining by Flow Cytometry

2.5 µg/10⁶ cells

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human/Mouse/Rat TRAM/TICAM2 by Western Blot. Western blot shows lysates of Raji human Burkitt's lymphoma cell line, C2C12 mouse myoblast cell line, and NRK rat normal kidney cell line. PVDF membrane was probed with 1 µg/mL of Human/Mouse/Rat TRAM/TICAM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4348) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for TRAM/TICAM2 at approximately 31 kDa (as indicated). This experiment was conducted using Immunoblot Buffer Group 2.

Immunocytochemistry

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TRAM/TICAM2 in Raji Human Cell Line. TRAM/TICAM2 was detected in immersion fixed Raji human Burkitt's lymphoma cell line using Human/Mouse/Rat TRAM/TICAM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4348) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, upper panel; Catalog # NL001) and counterstained with DAPI (blue, lower panel). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Immunocytochemistry

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TRAM/TICAM2 in C2C12 Mouse Cell Line. TRAM/TICAM2 was detected in immersion fixed C2C12 mouse myoblast cell line using Goat Anti-Human/Mouse/Rat TRAM/TICAM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4348) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, upper panel; Catalog # NL001) and counterstained with DAPI (blue, lower panel). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunocytochemistry

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TRAM/TICAM2 in NRK Rat Cell Line. TRAM/TICAM2 was detected in immersion fixed NRK rat normal kidney cell line using Goat Anti-Human/Mouse/Rat TRAM/TICAM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4348) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, upper panel; Catalog # NL001) and counterstained with DAPI (blue, lower panel). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Intracellular Staining by Flow Cytometry

Detection of TRAM/TICAM2 antibody in Raji Human Cell Line antibody by Flow Cytometry.

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Detection of TRAM/TICAM2 in Raji Human Cell Line by Flow Cytometry. Raji human Burkitt's lymphoma cell line was stained with Human/Mouse/Rat TRAM/TICAM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4348, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Intracellular Staining by Flow Cytometry

Detection of TRAM/TICAM2 antibody in C2C12 Mouse Cell Line antibody by Flow Cytometry.

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Detection of TRAM/TICAM2 in C2C12 Mouse Cell Line by Flow Cytometry. C2C12 mouse myoblast cell line was stained with Goat Anti-Human/Mouse/Rat TRAM/TICAM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4348, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Intracellular Staining by Flow Cytometry

Detection of TRAM/TICAM2 antibody in NRK Rat Cell Line antibody by Flow Cytometry.

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Detection of TRAM/TICAM2 in NRK Rat Cell Line by Flow Cytometry. NRK rat normal kidney cell line was stained with Goat Anti-Human/Mouse/Rat TRAM/TICAM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4348, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: TRAM/TICAM2

The innate and adaptive immune responses depend on systems that link cell surface surveillance receptor signals to cytoplasmic proteins such as kinases, adaptors, and transcription factors. Toll-like receptors (TLR) recognize different pathogen associated molecular patterns (PAMPs), and initiate a signaling cascades mediated by a Toll/interleukin-1 receptor (TIR) domain-containing adaptor proteins such as MyD88, TIRAP/MAL, and TRIF. Mouse TRIF-related adaptor molecule (TRAM), is a 232 amino acid, 26 kDa (predicted), ubiquitously expressed member of the TIR domain-containing adaptor family. TRAM, also known as TIR domain-containing adapter protein 2 (TICAM2) and TIR domain-containing protein (TIRP), contains a central Toll/interleukin-1 receptor (TIR) domain that is most similar to that of TRIF. TRAM plays an essential role in the MyD88-independent signaling of TLR4 by binding members of the IRAK family, ultimately leading to the activation of NF kappa B. Mouse TRAM shares 75% and 77% identity to human and rat TRAM, respectively.

Long Name

TRIF-related Adapter Molecule

Entrez Gene IDs

353376 (Human); 225471 (Mouse); 364867 (Rat)

Alternate Names

MGC129876; MGC129877; Putative NF-kappa-B-activating protein 502; TICAM2; TICAM-2TIR domain-containing adapter molecule 2; TIRAP3; TIRP; TIRPcytoplasmic adaptor; toll/interleukin-1 receptor (TIR) domain-containing adapter protein; Toll/interleukin-1 receptor domain-containing protein; toll-like receptor adaptor molecule 2; Toll-like receptor adaptor protein 3; TRAM; TRAMNF-kappa-B-activating protein 502; Trif; TRIF-related adapter molecule; TRIF-related adaptor molecule