This antibody should recognize Bcl-XL, Bcl-xS and Bcl-x(beta) as the immunogen sequence is common to them. The antibody does not cross-react with other Bcl-2 family members.
Synthetic peptide within Human Bcl-XL aa 1-100. The exact sequence is proprietary.
WB: Jurkat, K562, C6, RAW264.7, PC-12 NIH/3T3 cell lysates. IHC-P: Human endometrium and prostate carcinoma tissues. ICC/IF: HepG2, NIH/3T3, C6 and HeLa cells. Flow Cyt: DU145 and Jurkat cells. IP: Jurkat whole cell lysate (ab7899).
Our Abpromise guarantee covers the use of ab32370 in the following tested applications.
Potent inhibitor of cell death. Inhibits activation of caspases (By similarity). Appears to regulate cell death by blocking the voltage-dependent anion channnel (VDAC) by binding to it and preventing the release of the caspase activator, CYC1, from the mitochondrial membrane.
The BH4 motif is required for anti-apoptotic activity. The BH1 and BH2 motifs are required for both heterodimerization with other Bcl-2 family members and for repression of cell death.
Proteolytically cleaved by caspases during apoptosis. The cleaved protein, lacking the BH4 motif, has pro-apoptotic activity.
Mitochondrion membrane. Nucleus membrane. Mitochondrial membranes and perinuclear envelope.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human endometrium tissue labelling Bcl-XL with purified ab32370 at 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit HRP (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunocytochemistry/ Immunofluorescence analysis of C6(Rat glial tumor glial cell) cells labeling Bcl-XL with purified ab32370 at 1/100 dilution (1.42 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3(Mouse embryonic fibroblast) cells labeling Bcl-XL with purified ab32370 at 1/100 dilution (1.42 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
ab32370 (purified) at 1/500 dilution (0.28 µg/ml) immunoprecipitating Bcl-XL in NIH/3T3 whole cell lysate.
Flow cytometry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labelling Bcl-XL with ab32370 at 1/20 dilution (7.1 µg/ml) (Red). Cells were fixed with 4% paraformaldehyde . Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabeled control - Unlabelled cells (Blue).
Flow cytometry analysis of C6 (Rat glial tumor glial cell) cells labelling Bcl-XL with ab32370 at 1/20 dilution (7.1 µg/ml) (Red). Cells were fixed with 4% paraformaldehyde . Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabeled control - Unlabelled cells (Blue).
ICC/IF image of unpurified ab32370 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32370, 1/100) overnight at +4°C. The secondary antibody (green) was ab96899, goat anti-rabbit DyLight® 488 (IgG; H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Flow cytometry analysis of Jurkat cells labelling Bcl-XL with purified ab32370 at 1/20 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
ab32370 (purified) at 1/30 immunoprecipitating Bcl-XL in Jurkat cell lysate (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).
Western blot - Anti-Bcl-XL antibody [E18] (ab32370)Image courtesy of an anonymous Abreview.
Patient recieved anthracycline and taxane neoadjuvant chemotherapy.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-XL antibody [E18] (ab32370)Image from Medic S & Ziman M, PLoS One. 2010 Apr 22;5(4):e9977. Fig 5.; doi:10.1371/journal.pone.0009977; April 22, 2010, PLoS ONE 5(4): e9977.
Immunohistochemistry of human primary melanoma, staining Bcl-XL (red) with unpurified ab32370.
Antigen retrieval was performed in EDTA/Tris buffer (pH 8) before being blocked with 10%NGS for one hour at room temperature. Samples were incubated with primary antibody (1/50) at room temperature for one hour. An AlexaFluor®-conjugated anti-rabbit IgG was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling Bcl-XL with unpurified ab32370 at 1/50.
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Bcl-XL with purified ab32370 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, a goat anti-rabbit Alexa Fluor® 488 (IgG; 1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Overlay histogram showing DU145 cells stained with unpurified ab32370 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32370, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was a goat anti-rabbit DyLight® 488 (IgG; H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.