Human/Mouse/Rat TRAM/TICAM2 Antibody

  
  • 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 as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
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.
Data Examples
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
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
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
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 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 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 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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
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