Western blot abreview for Anti-Giantin antibody [9B6] - Golgi Marker

Inconclusive
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa, U2OS, 293T)
Loading amount
30 µg
Specification
HeLa, U2OS, 293T
Gel Running Conditions
Reduced Denaturing (5% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Other product details

Concentration
2 µg/ml
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 2.5% Milk, PBS-Tween 20

Secondary antibody

Name
Non-Abcam antibody was used: Goat anti-mouse HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000

Detection

Detection method
Supersignal west dura
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: >250 kDa ? kDa
Positive control
HeLa

Additional data

Additional Notes
Lane order (L-R): HeLa, U2OS, 293T
Full length giantin is approximately 370 kDa in size. This antibody failed to detect any obvious species of this Mw - although a series of high mw (>250 kDa) species were observed in all lysates, including HeLa - which express giantin as determined by ICC/IF. The upper range of the Mw markers is 250 kDa, therefore the mobility of these species is unclear. A strong band at ~200 kDa was observed in U2OS lysate, which is close to the predicted MW of a giantin transcript (variant c Aceview database).
Abcam response
This negative Abreview is currently being investigated by Abcam Technical Support. Please check here for an update response from Abcam.

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Submitted Jul 30 2008

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