Recombinant
RabMAb

Recombinant Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)

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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 - BSA and Azide free
  • Suitable for: ICC/IF, IP, Flow Cyt, IHC-P, WB
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Bcl-XL antibody [E18] - BSA and Azide free
    See all Bcl-XL primary antibodies

  • Description

    Rabbit monoclonal [E18] to Bcl-XL - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IP, Flow Cyt, IHC-P, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Pig

  • Immunogen

    Synthetic peptide within Human Bcl-XL aa 1-100. The exact sequence is proprietary.
    Database link: Q07817

  • Positive control

    • Jurkat whole cell lysate (ab7899) can be used as a positive control in WB.

  • General notes

    Ab199099 is the carrier-free version of ab32370. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab199099 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Applications

Our Abpromise guarantee covers the use of ab199099 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
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
WB Use at an assay dependent concentration. Predicted molecular weight: 26 kDa.

Please check the parent abID, ab32370, for more information on dilutions.

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

    see all

  • 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

Images

  • Western blot - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    Western blot - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099) + C6 (rat glioma) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

    Predicted band size: 26 kDa


    Exposure time: 30 seconds


    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM/TBST

  • Immunocytochemistry/ Immunofluorescence - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    Immunocytochemistry/ Immunofluorescence - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32370).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32370).

  • Flow Cytometry - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    Flow Cytometry - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32370).

  • Immunoprecipitation - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    Immunoprecipitation - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32370).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling Bcl-XL with unpurified ab32370 at 1/50.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32370).

  • Immunocytochemistry/ Immunofluorescence - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    Immunocytochemistry/ Immunofluorescence - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32370).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32370).

  • Flow Cytometry - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    Flow Cytometry - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32370).

  • Other - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    Other - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32370).

References (16)

Publishing research using ab199099? Please let us know so that we can cite the reference in this datasheet.

ab199099 has been referenced in 16 publications.

  • Bah N  et al. Bcl-xL controls a switch between cell death modes during mitotic arrest. Cell Death Dis 5:e1291 (2014). WB . PubMed: 24922075
  • Ho WT  et al. Dexamethasone modifies mitomycin C-triggered interleukin-8 secretion in isolated human Tenon's capsule fibroblasts. Exp Eye Res 124:86-92 (2014). Human . PubMed: 24858696
  • Wang B  et al. Role of Ku70 in deubiquitination of Mcl-1 and suppression of apoptosis. Cell Death Differ 21:1160-9 (2014). PubMed: 24769731
  • Redmond EM  et al. Perivascular delivery of Notch 1 siRNA inhibits injury-induced arterial remodeling. PLoS One 9:e84122 (2014). WB, ICC/IF ; Mouse . PubMed: 24416200
  • You S  et al. Disruption of STAT3 by niclosamide reverses radioresistance of human lung cancer. Mol Cancer Ther 13:606-16 (2014). PubMed: 24362463
View all Publications for this product

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