Overview

  • Product name
    Anti-Bcl-XL antibody [E18]
    See all Bcl-XL primary antibodies
  • Description
    Rabbit monoclonal [E18] to Bcl-XL
  • 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.
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Pig
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Bcl-XL aa 1-100.
    Database link: Q07817

  • 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 and HeLa cells. Flow Cyt: DU145 and Jurkat cells. IP: Jurkat cell lysate.
  • General notes

    A trial size is available to purchase for this antibody.

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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
WB 1/1000. Detects a band of approximately 26 kDa.
IHC-P 1/2000.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

ICC/IF 1/100 - 1/500.

 

Flow Cyt 1/20 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/10 - 1/30.

 

Target

  • 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
    modifications
    Proteolytically 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
  • Database links
  • Alternative names
    • Apoptosis regulator Bcl X antibody
    • Apoptosis regulator Bcl-X antibody
    • Apoptosis regulator BclX antibody
    • B cell lymphoma 2 like antibody
    • B2CL1_HUMAN antibody
    • Bcl 2 like 1 protein antibody
    • Bcl X antibody
    • Bcl xL antibody
    • BCL XL/S antibody
    • Bcl xS antibody
    • Bcl-2-like protein 1 antibody
    • Bcl2 Like 1 antibody
    • Bcl2 related gene antibody
    • Bcl2-L-1 antibody
    • BCL2L antibody
    • Bcl2l1 antibody
    • BCLX antibody
    • BclXL antibody
    • BclXs antibody
    • DKFZp781P2092 antibody
    • PPP1R52 antibody
    • Protein phosphatase 1 regulatory subunit 52 antibody
    see all

Anti-Bcl-XL antibody [E18] images

  • 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
    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.

  • 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.

  • 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.

  • 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
    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.

  • 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).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Anti-Bcl-XL antibody [E18] (ab32370) at 1/500 dilution (unpurified) + whole cell lysate prepared from a clinical sample of 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

    Image courtesy of an anonymous Abreview.

    Patient recieved anthracycline and taxane neoadjuvant chemotherapy.

    See Abreview

  • 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.

    Control: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

  • 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.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References for Anti-Bcl-XL antibody [E18] (ab32370)

This product has been referenced in:
  • Singh A  et al. Podophyllotoxin and Rutin Modulates Ionizing Radiation-Induced Oxidative Stress and Apoptotic Cell Death in Mice Bone Marrow and Spleen. Front Immunol 8:183 (2017). Read more (PubMed: 28289414) »
  • Yang MC  et al. Bim directly antagonizes Bcl-xl in doxorubicin-induced prostate cancer cell apoptosis independently of p53. Cell Cycle 15:394-402 (2016). WB, IP ; Human . Read more (PubMed: 26694174) »

See all 36 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Breast Cancer (clinical sample))
Loading amount
20 µg
Specification
Breast Cancer (clinical sample)
Treatment
patient recieved anthracycline and taxane neoadjuvant chemotherapy
Gel Running Conditions
Reduced Denaturing (12 %)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Submitted Jul 01 2011

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