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Western blot analysis of extracts from various cell lines using BOB-1/OBF-1 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, BOB-1/OBF-1 protein is not detected in C2C12 cells.Learn more about how we get our images.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with Myc/DDK-tagged hBOB-1/OFB-1 (+) using BOB-1/OBF-1 Antibody (upper), DYKDDDDK Tag Antibody #2368 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).Learn more about how we get our images.
Western blot analysis of extracts from various cell lines using BOB-1/OBF-1 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, BOB-1/OBF-1 protein is not detected in HDLM-2 cells.Learn more about how we get our images.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised October 2016
Protocol Id: 10
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
BOB-1/OBF-1 Antibody recognizes endogenous levels of total BOB-1/OBF-1 protein.
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro209 of human BOB-1/OBF-1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
B-cell Oct binding factor-1 (BOB-1/OBF-1) is a B-cell restricted transcriptional coactivator. BOB-1 facilitates transactivation of immunoglobulins and other B-cell specific genes through the binding and activation of the transcription factors Oct-1 and Oct-2 (1-4). Research studies have demonstrated that BOB-1 expression is required for antigen-dependent B-cell maturation (5-7). In pathological conditions such as classical Hodgkin’s disease, loss of BOB-1 expression is thought, in part, to contribute to the defect in immunoglobulin gene expression by Hodgkin and Reed Sternberg cells (8,9). In the context of multiple myeloma, overexpression of BOB-1 has been shown to contribute to malignant plasma cell cell growth, in part, through enhanced transactivation of TNFRSF17/BCMA (10).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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