Recombinant human Interferon gamma protein (Active) (ab9659)
Key features and details
- Expression system: Escherichia coli
- Purity: > 98% SDS-PAGE
- Endotoxin level:
- Active: Yes
- Suitable for: Functional Studies, SDS-PAGE, HPLC
Preparation and Storage
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Stability and Storage
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.
Constituent: PBS
This product is an active protein and may elicit a biological response in vivo, handle with caution.
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ReconstitutionCentrifuge vial prior to opening. Reconstitute in 100 µl 1x PBS, pH 8.0 to a concentration of 1.0 mg/ml. Do not vortex. Long term storage: Follow reconstitution with further dilution in a buffer containing a carrier protein (example; 0.1% BSA).
Images
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Western blot - Recombinant human Interferon gamma protein (ab9659)All lanes : Anti-CXCL11 antibody [EPR21755-173] (ab216157) at 1/1000 dilution
Lane 1 : THP-1 (hman monocytic leukemia cell line) whole cell lysate
Lane 2 : THP-1 treated with 200 ng/ml interferon-gamma (IFN-gamma, ab9659) and 50 ng/ml lipopolysaccharide (LPS) for 24 hours, then 300 ng/ml Brefeldin A (BFA) was added to the treated cells for 20 hours, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Observed band size: 11 kDa why is the actual band size different from the predicted?
Exposure time: 37 secondsBlocking/Dilution: 5% NFDM/TBST
The expression profile observed is consistent with what has been described in the literature (PMID: 17142784).
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Immunoprecipitation - Recombinant human Interferon gamma protein (ab9659)CXCL11 was immunoprecipitated from 0.35mg of THP-1 (human monocytic leukemia cell line) whole cell lysate with ab216157 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216157 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: THP-1 (human monocytic leukemia cell line) treated with 200 ng/ml interferon-gamma (IFN-gamma, ab9659) and 50 ng/ml lipopolysaccharide (LPS) for 24 hours, then added 300 ng/ml Brefeldin A (BFA) for 20 hours, whole cell lysate 10ug (Input).
Lane 2: ab216157 IP in THP-1 treated with 200 ng/ml interferon-gamma (IFN-gamma, ab9659) and 50 ng/ml lipopolysaccharide (LPS) for 24 hours, then added 300 ng/ml Brefeldin A (BFA) for 20 hours, whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216157 in THP-1 treated with 200 ng/ml interferon-gamma (IFN-gamma, ab9659) and 50 ng/ml lipopolysaccharide (LPS) for 24 hours, then added 300 ng/ml Brefeldin A (BFA) for 20 hours, whole cell lysate.Blocking/Dilution buffer: 5% NFDM/TBST
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Western blot - Recombinant human Interferon gamma protein (ab9659)All lanes : Anti-IP10 antibody [EPR20764] (ab214668) at 1/1000 dilution
Lane 1 : Untreated THP-1 (human monocytic leukemia cell line) culture supernatant
Lane 2 : THP-1 treated with 200 ng/ml interferon-gamma (IFN-gamma, ab9659) and 50 ng/ml lipopolysaccharides (LPS) for 24 hours, culture supernatant
Lysates/proteins at 15 µl per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Observed band size: 12 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
IP10 protein secretion can be induced by IFN-gamma treatment (PMID: 11907072).
