Mouse, Human Predicted to work with:
Horse, Dog, Pig, Macaque monkey, Gorilla
Synthetic peptide corresponding to Mouse Iba1 aa 1-100 conjugated to Keyhole Limpet Haemocyanin (KLH). (Peptide available as ab125894)
Human hippocampus formalin fixed paraffin embedded tissue section; Human and mouse spleen tissue lysate; U937, MOLT4, THP1, Raw 264.7, NR8383 whole cell lysates
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 16 kDa).
For WB we recommend blocking in 3% milk. Blocking with BSA gives high background.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Actin-binding protein that enhances membrane ruffling and RAC activation. Enhances the actin-bundling activity of LCP1. Binds calcium. Plays a role in RAC signaling and in phagocytosis. May play a role in macrophage activation and function. Promotes the proliferation of vascular smooth muscle cells and of T-lymphocytes. Enhances lymphocyte migration. Plays a role in vascular inflammation.
Detected in T-lymphocytes and peripheral blood mononuclear cells.
Contains 2 EF-hand domains.
Phosphorylated on serine residues.
Cytoplasm > cytoskeleton. Cell projection > ruffle membrane. Associated with the actin cytoskeleton at membrane ruffles and at sites of phagocytosis.
All lanes : Anti-Iba1 antibody (ab105464) at 5 µg/ml
Lane 1 : HEK293 whole cell lysate at 20 µg Lane 2 : A431 whole cell lysate at 20 µg Lane 3 : Human spleen tissue lysate at 30 µg Lane 4 : Mouse spleen tissue lysate at 30 µg Lane 5 : U937 whole cell lysate at 30 µg Lane 6 : MOLT4 whole cell lysate at 30 µg Lane 7 : THP1 whole cell lysate at 30 µg Lane 8 : THP1 whole cell lysate, PMA treated at 30 µg Lane 9 : Raw 264.7 whole cell lysate at 30 µg Lane 10 : C6 whole cell lysate at 30 µg Lane 11 : NR8383 whole cell lysate at 30 µg
All lanes blocked using 3% milk. For ab105464 in WB we recommend blocking in milk. Blocking with BSA gives high background.
This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with ab108539 (anti-Iba1 antibody; 1/1000) for 18 hours at 4°C. Antibody binding was detected using HRP-labelled anti-Rabbit IgG for 1 hour at room temperature and visualised using ECL development solution ab133406.
IHC image of ab105464 staining in human hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH9, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab105464, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times