Immunofluorescence analysis of Bcl-X Antibody was done on 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Bcl-X Antibody (AHO0222) at 1µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour 488 Rabbit Anti-Mouse IgG Secondary Antibody (A11059) at a dilution of 1:400 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing cytoplasmic and nuclear localization. Panel e is a no primary antibody control. The images were captured at 40X magnification.
|Tested species reactivity||Human, Mouse, Non-human primate, Pig, Rat|
|Published species reactivity||Human, Not Applicable|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||A synthetic peptide, aa 3-14 (Cys-QSNRELVVDFLS), of human Bcl-X protein. Thisamino acid sequence is shared by human and murine Bcl-X protein.|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||Assay Dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||Assay-Dependent|
|Western Blot (WB)||1:500-1:2000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody reacts with both Bcl-XS (short) and Bcl-XL (long) proteins. The immunogenic sequence (aa 3-14) is conserved in human and mice.
Staining of formalin-fixed paraffin-embedded tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0, for 20 min followed by cooling at room temperature for 20 min. A recommended positive control is Hodgkin's lymphoma.
Bcl-X has two isoforms, Bcl-XL (long), a 241 amino acid protein, and Bcl-XS (short), a 178 amino acid protein lacking a 63 amino acid domain that is well-conserved among members of the Bcl-2 family. This family of proteins is an important regulator of apoptosis. Bcl xL forms heterodimers with BAX, BAK, and Bcl2, and its overexpression in tumor cells confers resistance against chemotherapeutic drugs. Bcl xL is phosphorylated on many sites including serine 62, a critical site for Bcl xL response to microtubule-damaging drugs such as taxol and vinblastine. Phosphorylation of serine 62 is thought to be mediated by Jun N-terminal stress kinase (JNK) signaling - negatively regulates the anti-apoptotic function of Bcl xL and controls the growth of neoplastic cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Human||1:50||Immunohistochemical markers of tissue injury in biopsies with transplant glomerulitis.||Batal I,Azzi J,El-Haddad N,Riella LV,Lunz JG,Zeevi A,Sasatomi E,Basu A,Tan H,Shapiro R,Randhawa P||Human pathology (43:69)||2012|
|Human||Not Cited||Clinical and immunohistochemical features associated with a response to bortezomib in patients with multiple myeloma.||Dawson MA,Opat SS,Taouk Y,Donovan M,Zammit M,Monaghan K,Horvath N,Roberts AW,Prince HM,Hertzberg M,McLean CA,Spencer A||Clinical cancer research : an official journal of the American Association for Cancer Research (15:714)||2009|
Beclin 1-independent autophagy induced by a Bcl-XL/Bcl-2 targeting compound, Z18.
AHO0222 was used in western blot to identify and characterize a new compound that targets Bcl-XL/Bcl-2
|Tian S,Lin J,Jun Zhou J,Wang X,Li Y,Ren X,Yu W,Zhong W,Xiao J,Sheng F,Chen Y,Jin C,Li S,Zheng Z,Xia B||Autophagy (6:1032)||2010|
Differential expression of bcl-2 family proteins in bladder carcinomas. Relationship with apoptotic rate and survival.
AHO0222 was used in immunohistochemistry to study the role of various bcl-2 family molecules in the regulation of apoptosis and the progression of urothelial cancer.
|Korkolopoulou P,Lazaris ACh,Konstantinidou AE,Kavantzas N,Patsouris E,Christodoulou P,Thomas-Tsagli E,Davaris P||European urology (41:274)||2002|
|Not Applicable||Not Cited||
Molecular prognosticators of complex karyotype soft tissue sarcoma outcome: a tissue microarray-based study.
AHO0222 was used in immunohistochemistry - paraffin section to identify prognostic biomarkers for complex karyotype soft tissue sarcoma
|Lahat G,Tuvin D,Wei C,Wang WL,Pollock RE,Anaya DA,Bekele BN,Corely L,Lazar AJ,Pisters PW,Lev D||Annals of oncology : official journal of the European Society for Medical Oncology (21:1112)||2010|
Bcl-x(S) induces an NGF-inhibitable cytochrome c release.
AHO0222 was used in immunocytochemistry to elucidate the role of Bcl-x(S) in nerve growth factor-inhibitable apoptosis
|Lindenboim L,Schlipf S,Kaufmann T,Borner C,Stein R||Experimental cell research (297:392)||2004|