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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG_2B

Scientific Data Images for Human/Mouse/Rat GIT1 Antibody

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.

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.

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).

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Applications for Human/Mouse/Rat GIT1 Antibody

Application

Recommended Usage

CyTOF-ready

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

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
Sample: Immersion fixed paraffin-embedded sections of human brain (hippocampus)

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells
Sample: SH‑SY5Y human neuroblastoma cell line fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005)

Western Blot

2 µg/mL
Sample: 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

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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: 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

Alternate Names

ARF GAP GIT1

Entrez Gene IDs

28964 (Human); 216963 (Mouse); 83709 (Rat)

Gene Symbol

GIT1

UniProt

Q9Y2X7

Additional GIT1 Products

Product Documents for Human/Mouse/Rat GIT1 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human/Mouse/Rat GIT1 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human/Mouse/Rat GIT1 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB8508

Key Product Details

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Product Specifications

Immunogen

Specificity