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9934
Bcl-2 Family Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Bcl-2 Family Antibody Sampler Kit #9934

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Simple Western™ analysis of lysates (1.0 mg/mL) from HT-1080 cells using Bax Antibody #2772. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western blot analysis of extracts from Jurkat (human), NIH/3T3 (mouse) or C6 (rat) cells, untreated or etoposide-treated (25 µM, Jurkat) or staurosporine-treated (1 µM, NIH/3T3 and C6), using Bcl-xL Antibody.
Western blot analysis of extracts from control HeLa cells (lane 1) or Bax knockout HeLa cells (lane 2) using Bax Antibody #2772 (upper), or β-actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the Bax-knockout HeLa cells confirms specificity of the antibody for Bax.
Western blot analysis of extracts from 293, A431 and SK-N-MC (human) cells and COS (monkey) cells, using Bak Antibody.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Bad siRNA I (+), using Bad Antibody and p42 MAPK Antibody #9108. The Bad antibody confirms silencing of Bad expression, while the p42 MAPK antibody is used to control for loading and specificity of Bad siRNA.
Western blot analysis of extracts from COS cells untreated or TPA-treated, using Phospho-Bad (Ser112) (7E11) Mouse mAb (upper) or Bad Antibody #9292 (lower).
Immunoprecipitation of Bcl-xL from Jurkat cell extracts, using Bcl-xL Antibody. Lane 1 is the lysate control; lane 2 is antibody alone and lane 3 is antibody plus lysate.
Western blot analysis of extracts from 293 (human), COS (monkey), L929 (mouse) and PC12 (rat) cells, using Bax Antibody.
Immunoprecipitation of Bak from HeLa cell extracts, using Bak Antibody. Lane 1 is antibody alone, lane 2 is the lysate with protein A beads (negative control) and lane 3 is antibody plus lysate.
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Bad siRNA II (+), using Bad Antibody and α-Tubulin (11H10) Rabbit mAb #2125. The Bad antibody confirms silencing of Bad expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Bad siRNA.
Western blot analysis of extracts from 293 cells transfected with Wild-type Bad, Bad (Ser112A), Bad (S136A), Bad (S112A/S136A) and treated with TPA or forskolin, using Phospho-Bad (Ser112) (7E11) Mouse mAb (upper) or Bad Antibody #9292 (lower).
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-), SignalSilence® Bax siRNA I (+) or SignalSilence® Bax siRNA II (+), using Bax Antibody and α-Tubulin (11H10) Rabbit mAb #2125. The Bax Antibody confirms silencing of Bax expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Bax siRNA.
Western blot analysis of extracts from COS cells, untreated or TPA-treated, using Bad Antibody (right) or Phospho-Bad (Ser112) Antibody #9291 (left).
Immunoprecipitation of Bax from Jurkat cell extract, using Bax Antibody. Lane 1 is the lysate control; lane 2 is antibody alone; lane 3 is antibody plus lysate.
Western blot analysis of extracts from 293 cells transfected with Wild-type Bad, Bad (Ser112A), Bad (S136A) or Bad (S112A/S136A), untreated, TPA-treated or forskolin-treated, using Phospho-Bad (Ser112) Antibody #9291 (top), Phospho-Bad (Ser136) Antibody #9295 (middle) or Bad Antibody (bottom).
Inquiry Info.# 9934

Specificity / Sensitivity

Each antibody in the Bcl-2 Family Antibody Sampler Kit recognizes only its specific target. The antibodies do not cross-react with one another.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to the residues surrounding Ser70 of human Bcl-2; Asp61 of human Bcl-xL; the amino-terminal residues of human Bax and Bak; and Ser112 of mouse Bad. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the residues surrounding Ser112 of mouse Bad. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

Bcl-2 exerts a survival function in response to a wide range of apoptotic stimuli through inhibition of mitochondrial cytochrome c release (1). It has been implicated in modulating mitochondrial calcium homeostasis and proton flux (2). Several phosphorylation sites have been identified within Bcl-2, including Thr56, Ser70, Thr74, and Ser87 (3). It has been suggested that these phosphorylation sites may be targets of the ASK1/MKK7/JNK1 pathway and that phosphorylation of Bcl-2 may be a marker for mitotic events (4,5). Mutation of Bcl-2 at Thr56 or Ser87 inhibits its anti-apoptotic activity during glucocorticoid-induced apoptosis of T lymphocytes (6). Interleukin-3 and JNK-induced Bcl-2 phosphorylation at Ser70 may be required for its enhanced anti-apoptotic functions (7).

Description: The Bcl-2 Family Antibody Sampler Kit contains enough primary and secondary antibodies to perform four Western mini-blot experiments.

Pathways

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Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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