AIF-1/Iba1 Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBL1-07408
Key Product Details
Species
Human
Applications
Western Blot
Product Summary for AIF-1/Iba1 Overexpression Lysate
AIF-1/Iba1 Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC203154
Accession#: NM_032955
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Expression Host: HEK293T
Plasmid: RC203154
Accession#: NM_032955
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Product Specifications
Application Notes
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
TMW
16.7 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Type
Overexpression
Scientific Data Images for AIF-1/Iba1 Overexpression Lysate
![Western Blot: AIF-1/Iba1 Overexpression Lysate [NBL1-07408]](https://resources.bio-techne.com/images/products/Iba1-Overexpression-Lysate-Adult-Normal-Western-Blot-NBL1-07408-img0002.jpg "Western Blot: AIF-1/Iba1 Overexpression Lysate [NBL1-07408]")
Western Blot: AIF-1/Iba1 Overexpression Lysate [NBL1-07408]
Western Blot: AIF-1/Iba1 Overexpression Lysate [NBL1-07408] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right - AIF-1/Iba1 Overexpression LysateFormulation, Preparation, and Storage
Formulation
RIPA buffer
Concentration
The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.
Shipping
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store at -80C. Avoid freeze-thaw cycles.
Background: AIF-1/Iba1
Several cellular functions have been associated with AIF-1/Iba1 expression including cell growth, cell migration, actin bundling, membrane ruffling, and phagocytic activity (2, 4). Iba1 induces Rac signaling through a PLC-gamma dependent pathway (1). Rac, a member of the Rho family of small GTPases, localizes with Iba1 and F-actin in membrane ruffles and phagocytic cups and plays a role in microglia activation. AIF-1/Iba1 induction in macrophages and microglia occur in association with immunological inflammatory processes in various disease states including endometriosis, cerebral infarction and rheumatoid arthritis (5). Immunodetection of Iba1 through flow cytometry, immunohistochemical or immunocytochemical applications is commonly used for identification and analysis of microglia.
References
1. Imai, Y., & Kohsaka, S. (2002). Intracellular signaling in M-CSF-induced microglia activation: Role of Iba1. GLIA. https://doi.org/10.1002/glia.10149
2. Deininger, M. H., Meyermann, R., & Schluesener, H. J. (2002). The allograft inflammatory factor-1 family of proteins. FEBS Letters. https://doi.org/10.1016/S0014-5793(02)02430-4
3. Utans, U., Quist, W. C., Mcmanus, B. M., Wilson, J. E., Arceci, R. J., Wallace, A. F., & Russell, M. E. (1996). Allograft inflammatory factor-1: A cytokine-responsive macrophage molecule expressed in transplanted human hearts. Transplantation. https://doi.org/10.1097/00007890-199605150-00018
4. Franco-Bocanegra, McAuley, Nicoll, & Boche. (2019). Molecular Mechanisms of Microglial Motility: Changes in Ageing and Alzheimer's Disease. Cells. https://doi.org/10.3390/cells8060639
5. Kimura, M., Kawahito, Y., Obayashi, H., Ohta, M., Hara, H., Adachi, T., ... Yoshikawa, T. (2007). A Critical Role for Allograft Inflammatory Factor-1 in the Pathogenesis of Rheumatoid Arthritis. The Journal of Immunology. https://doi.org/10.4049/jimmunol.178.5.3316
Long Name
Allograft Inflammatory Factor 1
Alternate Names
AIF1, IBA1, IRT1
Gene Symbol
AIF1
Additional AIF-1/Iba1 Products
Product Documents for AIF-1/Iba1 Overexpression Lysate
Product Specific Notices for AIF-1/Iba1 Overexpression Lysate
HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.
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