Recombinant Anti-Bcl-XL antibody [E18] (ab32370)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E18] to Bcl-XL
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Bcl-XL antibody [E18]
See all Bcl-XL primary antibodies -
Description
Rabbit monoclonal [E18] to Bcl-XL -
Host species
Rabbit -
Specificity
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.
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Tested applications
Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Pig -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- 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 (intra): DU145 and Jurkat cells. IP: Jurkat whole cell lysate (ab7899).
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Dissociation constant (KD)
KD = 6.50 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E18 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32370 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (8) |
1/1000. Detects a band of approximately 26 kDa.
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/100 - 1/500.
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IP |
1/10 - 1/30.
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Flow Cyt (Intra) |
1/20 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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WB
1/1000. Detects a band of approximately 26 kDa. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/500.
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IP
1/10 - 1/30.
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Flow Cyt (Intra)
1/20 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Function
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.
Isoform Bcl-X(S) promotes apoptosis. -
Tissue specificity
Bcl-X(S) is expressed at high levels in cells that undergo a high rate of turnover, such as developing lymphocytes. In contrast, Bcl-X(L) is found in tissues containing long-lived postmitotic cells, such as adult brain. -
Sequence similarities
Belongs to the Bcl-2 family. -
Domain
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. -
Post-translational
modificationsProteolytically cleaved by caspases during apoptosis. The cleaved protein, lacking the BH4 motif, has pro-apoptotic activity. -
Cellular localization
Mitochondrion membrane. Nucleus membrane. Mitochondrial membranes and perinuclear envelope. - Information by UniProt
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Database links
- Entrez Gene: 598 Human
- Entrez Gene: 12048 Mouse
- Entrez Gene: 397536 Pig
- Entrez Gene: 24888 Rat
- Omim: 600039 Human
- SwissProt: Q07817 Human
- SwissProt: Q64373 Mouse
- SwissProt: O77737 Pig
see all -
Alternative names
- Apoptosis regulator Bcl X antibody
- Apoptosis regulator Bcl-X antibody
- Apoptosis regulator BclX antibody
see all
Images
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All lanes : Anti-Bcl-XL antibody [E18] (ab32370) at 1/1000 dilution (purified)
Lane 1 : Jurkat cell lysate
Lane 2 : K562 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Observed band size: 26 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-XL antibody [E18] (ab32370)
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.
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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.
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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.
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ab32370 (purified) at 1/20 dilution immunoprecipitating Bcl-XL in NIH/3T3 whole cell lysate.
Lane 1 (input): NIH/3T3(Mouse embryonic fibroblast) whole cell lysate 10µg
Lane 2 (+): ab32370 & NIH/3T3 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32370 in NIH/3T3 whole cell lysate
For western blotting, ab32370 at 1/500 dilution (0.28 µg/ml)
and VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM /TBST . -
Intracellular 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).
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Intracellular 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).
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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.
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All lanes : Anti-Bcl-XL antibody [E18] (ab32370) at 1/1000 dilution (purified)
Lane 1 : C6 cell lysate
Lane 2 : RAW264.7 cell lysate
Lane 3 : PC-12 cell lysate
Lane 4 : NIH/3T3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Observed band size: 26 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Intracellular 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.
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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).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-Bcl-XL antibody [E18] (ab32370) at 1/500 dilution (unpurified) + whole cell lysate prepared from a clinical sample of human breast cancer cells at 20 µg
Secondary
Goat anti-rabbit IgG-HRP at 1/1000 dilution
Developed using the ECL technique.
Observed band size: 26 kDa why is the actual band size different from the predicted?
Exposure time: 15 minutes
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) - Anti-Bcl-XL antibody [E18] (ab32370)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling Bcl-XL with unpurified ab32370 at 1/50.
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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.
Control: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
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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 22°C. The secondary antibody used was a goat anti-rabbit DyLight® 488 (IgG; H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (178)
ab32370 has been referenced in 178 publications.
- Xiao Z et al. Pimozide augments bromocriptine lethality in prolactinoma cells and in a xenograft model via the STAT5/cyclin D1 and STAT5/Bcl-xL signaling pathways. Int J Mol Med 47:113-124 (2021). PubMed: 33155660
- Ma Y et al. SphK1 promotes development of non-small cell lung cancer through activation of STAT3. Int J Mol Med 47:374-386 (2021). PubMed: 33236138
- Ren Y et al. Alternol Sensitizes Renal Carcinoma Cells to TRAIL-Induced Apoptosis. Front Pharmacol 12:560903 (2021). PubMed: 33841136
- Liang X & Ju J Matrine inhibits ovarian cancer cell viability and promotes apoptosis by regulating the ERK/JNK signaling pathway via p38MAPK. Oncol Rep 45:N/A (2021). PubMed: 33786627
- Qu Y et al. The neuroprotection of deproteinized calf blood extractives injection against Alzheimer's disease via regulation of Nrf-2 signaling. Aging (Albany NY) 13:11150-11169 (2021). PubMed: 33819182