Human/Mouse/Rat GIT1 Antibody

    
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  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human GIT1 in direct ELISAs and human, mouse, and rat GIT1 in Western blots. In sandwich immunoassays, this antibody is specific for human GIT1 when paired with the suggested detection antibody.
  • Source
    Monoclonal Mouse IgG2B Clone # 924640
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human GIT1
    Ser485-Asp636
    Accession # Q9Y2X7
  • 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
Product Datasheets

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Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
  • ELISA

    This antibody functions as an ELISA capture antibody when paired with Mouse Anti-Human GIT1 Monoclonal Antibody (Catalog # MAB85081). In sandwich immunoassays, this antibody is specific for human GIT1 when paired with the suggested detection antibody.

    This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human GIT1 DuoSet ELISA Kit (Catalog # DY8485-05) for convenient development of a sandwich ELISA.

  • Immunohistochemistry
    8-25 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.
  • Intracellular Staining by Flow Cytometry
    0.25 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, and Rat GIT1 by Western Blot. Western blot shows lysates of HUVEC human umbilical vein endothelial cells, U2OS human osteosarcoma cell line, M1 mouse myeloid leukemia cell line, and Rat‑2 rat embryonic fibroblast cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human GIT1 Monoclonal Antibody (Catalog # MAB8508) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for GIT1 at approximately 95 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunohistochemistry
GIT1 in Human Brain. GIT1 was detected in immersion fixed paraffin-embedded sections of human brain (hippocampus) using Mouse Anti-Human GIT1 Monoclonal Antibody (Catalog # MAB8508) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Intracellular Staining by Flow Cytometry
Detection of GIT1 in SH‑SY5Y Human Cell line by Flow Cytometry. SH‑SY5Y human neuroblastoma cell line was stained with Mouse Anti-Human GIT1 Monoclonal Antibody (Catalog # MAB8508, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).
Human GIT1 ELISA Standard Curve. Recombinant Human GIT1 protein was serially diluted 2-fold and captured by Mouse Anti-Human/Mouse/Rat GIT1 Monoclonal Antibody (Catalog # MAB8508) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-Human GIT1 Monoclonal Antibody (Catalog # MAB85081) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
    Reconstitution Buffer Available
  • 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: GIT1
GIT1 (ARF GTPase-activating protein GIT1) is a 95 kDa protein that belongs to ADP ribosylation factor family and is localized to focal adhesions, cytoplasmic complexes and membrane protrusions, and regulates cell protrusion formation and cell migration. G-protein coupled receptor (GPCR) kinase interacting proteins 1 and 2 (GIT1 and GIT2) are highly conserved, ubiquitous scaffold proteins involved in localized signaling to help regulate focal contact assembly and cytoskeletal dynamics. GIT proteins contain multiple interaction domains that allow interaction with small GTPases (including ARF, Rac and cdc42), kinases (such as PAK and MEK), the Rho family GEF PIX, and the focal adhesion protein paxillin. GIT1 has also been implicated in neuronal functions including synapse formation and the pathology of Huntington disease. Huntington disease is a genetic neurodegenerative condition involving a mutation in the huntington gene. The huntington gene product (htt) is ubiquitinated and degraded in human Huntington disease brains. Htt interacts directly with GIT1 causing enhanced htt proteolysis, indicating that GIT1 distribution and function may contribute to Huntington disease pathology. Within amino acids (aa) 485-636, human and mouse GIT1 share 93% aa sequence identity.
  • Long Name:
    G Protein-Coupled Receptor Kinase Interacting ArfGAP 1
  • Entrez Gene IDs:
    28964 (Human); 216963 (Mouse); 83709 (Rat)
  • Alternate Names:
    ARF GAP GIT1; ARF GTPase-activating protein GIT1; CAT1; CAT-1; Cool-associated and tyrosine-phosphorylated protein 1; G protein-coupled receptor kinase interacting ArfGAP 1; G protein-coupled receptor kinase interactor 1; G protein-coupled receptor kinase-interactor 1; GIT1; GRK-interacting protein 1
Related Research Areas

FAQs

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