BAX antibody - N-terminal region (ARP58870_P050)

Catalog#: ARP58870_P050

• Size: 50ug

Human HepG2

• Description of Target: The protein encoded by this gene belongs to the BCL2 protein family. BCL2 family members form hetero- or homodimers and act as anti- or pro-apoptotic regulators that are involved in a wide variety of cellular activities. This protein forms a heterodimer with BCL2, and functions as an apoptotic activator. This protein is reported to interact with, and increase the opening of, the mitochondrial voltage-dependent anion channel (VDAC), which leads to the loss in membrane potential and the release of cytochrome c. The expression of this gene is regulated by the tumor suppressor P53 and has been shown to be involved in P53-mediated apoptosis. Multiple alternatively spliced transcript variants, which encode different isoforms, have been reported for this gene.
• Gene Symbol: BAX
• Official Gene Full Name: BCL2-associated X protein
• Alias Symbols: BCL2L4
• Protein Accession#: NP_004315
• Nucleotide Accession#: NM_004324
• Swissprot Id: Q07812
• Host: Rabbit
• Clonality: Polyclonal
• Protein Size (# AA): 218
• Molecular Weight: 24kDa
• Application: WB
• Partner Proteins: TP53, TP53, TP73, BBC3, BCL2, BCL2, BCL2L1, BCL2L1, BID, MOAP1, NOL3, BAK1, BBC3, BCL2, BCL2A1, BCL2L1, BCL2L10, BID, ERN1, KCNA3, MCL1, MOAP1, NOL3, PMAIP1, PPP1CA, SFN, SH3GLB1, SLC25A4, VDAC1, YWHAQ, GTF2B, MUC1, TAF1, TBP, TP53, BAK1, BAX, BCL2, BCL2A1, BCL2L1, BCL2L1, BCL2L10, BCL2L2, BNIPL, Bak1, CRYAA, CRYAB, HSPD1, MCL1, MEPCE, MOAP1, MOAP1, OYE2, PMAIP1, SH3GLB1, SH3GLB1, SLC25A4, SNCA, TOMM22, TOMM40, UBC, VDAC1, YWHAQ
• Immunogen: The immunogen for anti-BAX antibody: synthetic peptide directed towards the N terminal of human BAX
• Product Format: Lyophilized powder
• Purification: Affinity Purified
• Predicted Homology Based on Immunogen Sequence: Bovine: 100%; Dog: 100%; Horse: 100%; Human: 100%; Mouse: 100%; Pig: 100%; Rat: 100%; Sheep: 100%; Rabbit: 90%
• Peptide Sequence: Synthetic peptide located within the following region: IMKTGALLLQGFIQDRAGRMGGEAPELALDPVPQDASTKKLSECLKRIGD
• Blocking Peptide: For anti-BAX antibody is Catalog # AAP58870 (Previous Catalog # AAPP44817)
• Key Reference: N/A
• Reconstitution and Storage: Add 50 μl of distilled water. Final anti-BAX antibody concentration is 1 mg/ml in PBS buffer. For longer periods of storage, store at -20°C. Avoid repeat freeze-thaw cycles.

Product Protocols: BAX antibody tested with Human Hepg2 Cells (ARP58870_P050)

Aviva Systems Biology is the original manufacturer of this BAX antibody (ARP58870_P050)

Click here to view the BAX antibody Western Blot Protocol

Product Datasheet Link: BAX antibody (ARP58870_P050)

WB Suggested Anti-BAX Antibody Titration: 0.2-1 ug/ml
ELISA Titer: 1:312500
Positive Control: HepG2

Western Blot image:


Description of Target: The protein encoded by this gene belongs to the BCL2 protein family. BCL2 family members form hetero- or homodimers and act as anti- or pro-apoptotic regulators that are involved in a wide variety of cellular activities. This protein forms a heterodimer with BCL2, and functions as an apoptotic activator. This protein is reported to interact with, and increase the opening of, the mitochondrial voltage-dependent anion channel (VDAC), which leads to the loss in membrane potential and the release of cytochrome c. The expression of this gene is regulated by the tumor suppressor P53 and has been shown to be involved in P53-mediated apoptosis. Multiple alternatively spliced transcript variants, which encode different isoforms, have been reported for this gene.

Questions pertaining to this data can be directed to techsupport@avivasysbio.com

Aviva Systems Biology’s BAX antibody (ARP58870_P050) has been tested using other cell lysates and tissues. To obtain more data about this antibody please email us at info@avivasysbio.com.

To order by phone call us at (888) 880-0001, fax us at (858) 552-6975 or send an email to info@avivasysbio.com. Aviva manufactures this antibody so we can offer the best price. Please contact us to request pricing information.

All of Aviva’s products are guaranteed for the applications and experimental sample types mentioned in the datasheet below. Are you curious if this product will work for you? Please contact us at techsupport@avivasysbio.com

Product Review: BAX antibody – N-terminal region (ARP58870_P050) in xenopus laevis embryo using Western Blot

Product Page for BAX antibody - N-terminal region (ARP58870_P050)

Researcher: Rosa Carotenuto, Department of Structural and Functional Biology, University of Naples 'Federico II'
Application: Western blotting
Species+tissue/cell type: xenopus laevis embryos
How many ug’s of tissue/cell lysate run on the gel:  1. 50 ug xenopus laevis embryos lysate (lysate buffer) 2. 50 ug xenopus laevis embryos lysate (HB buffer)
Primary antibody dilution: 1:400
Secondary antibody: Goat Anti-Rabbit AP
Secondary antibody dilution: 1:20000

Questionnaire:

How do Aviva’s reagents play a role in your experimental goals?

Actually not

Questionnaire:

How would you rate this antibody on a scale from 1-5 (5=best) and why?

5, because it is specific

Would you use this antibody in future experiments?

Yes.

Have you used another antibody which has worked in your application?

Yes.

Do you believe the information about the reagent on Aviva’s website is correct?

Yes.

If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?

Yes,because the binding specificity appears to be appreciable.

Sample Description (please include 1) species type, and 2) cell/tissue type, and 3) how much protein loaded for each lane of your gel):

1)Xenopus laevis; 2) total homogenate; 3) 50micrograms

How many different experimental trials were conducted using the antibody sample?

Two trials of protein extraction utilising HB (25mM Hepes, pH7.5, containing 900mM glycerol, 0.02mM NaN3, 1mM ATP, 1mM DTT, 5mM EGTA) and lisys buffer(TRIS 20mM pH7.5, NaCl 140mM, glicerolo 10%, DTT 1mM, sodium vanadate 2mM, NaF 25mM, Nonidet 1%) and two trials for immunoblotting.

What type of experimental sample are you using and how did you preparing it?

Embryos were grouped according to their stage and homogenized in ice-cold 25mM Hepes, pH7.5, containing 900mM glycerol, 0.02mM NaN3, 1mM ATP, 1mM DTT, 5mM EGTA (HB) or in the Lisys buffer (TRIS 20mM pH7.5, NaCl 140mM, glicerolo 10%, DTT 1mM, sodium vanadate 2mM, NaF 25mM, Nonidet 1%) and the following protease inhibitors (Sigma): 2mMTAME, 5mg/mlSBTI, 5 mg/ml aprotinin and 10 mM E64. Aftercentrifugation at 15,000g for 40 min at 4 °C, the supernatants were boiled for 3 min in sample buffer containing 50mM Tris-HCl pH 6.8, 69mM SDS, 0.028 mM bromophenol blue, 1 mM DTT. Aliquots of protein were submitted to SDS-PAGE using protein standards as reference markers (200,116,97,45,31and21kDa)(Bio-Rad, Hercules,CA,USA). Sixty mg of proteins from embryos of all the embryogenetic stages were loaded on each gel. The samples were transferred electrophoretically on nitrocellulose paper. Antibody binding was detected by goat anti- rabbit IgG conjugated to alkaline phosphatise and revealed using the chromogenic substrates NBT and  BCIP.

Primary used and dilution:

1:400 for 1 hour at room temperature.

Secondary used and dilution:

Antirabbit IgG APconjugated 1:20.000 for 1 hour at room temperature.

Experimental Procedure/Protocols:

Western blotting was performed on nitrocellulose membrane (NC) Protran (Schleicher and Schuell). The gel and the membrane have been brought into contact and then covered on both sides of sheets of filter paper and sponges, previously impregnated by the transfer solution (0.02M Tris, 0.13M glycine and methanol in varying proportions depending on the molecular weight of the protein to be transferred). The transfer was carried out for 2h at 400mA at 4ºC. The transfer efficiency was evaluated by staining the nitrocellulose filter with Ponceau S (Sigma). The gel is stained with the corresponding 0.1% / 20% ethanol Coomassie Brilliant Blue (Biorad). The NC was incubated with 3% BSA (bovine serum albumin, SIGMA) in TBS (Tris 10mM, NaCl 154mm and H2O, pH 7.4) 2h at 37°C, to block nonspecific binding sites. After extensive washes in TBS, the NC was incubated with primary antibody, diluted in 3% BSA / 1% Triton X100/TBS, two hours at RT or ON at 4°C depending on the antibody used. Washes in 0.1% Triton X100/TBS. The secondary antibody (anti-mouse or anti-rabbit-AP conjugate) diluted in 0.1% Triton X100/TBS, was incubated for 1h at RT. Abundant washes in 0.1% Triton X100/TBS. Incubation in AP buffer (100 mM Tris, 100 mM NaCl, 5 mM MgCl2, pH 9.5. Revelation using the Chromogenic substrates NBT and BCIP. Stop in 20 mM Tris, 15 mm EDTA, pH 8.

What controls were used in your experiment? Please include your positive control:

Negative controls were used by omitting the primary antibody or by replacing the primary antibody with rabbit IgG.

How did you store the antibody after re-suspension?

We store the antibody at -20°C (stock) or at 4°C (in use).