Overview

  • Product name
    Anti-Bcl-XL antibody [EPR16642]
    See all Bcl-XL primary antibodies
  • Description
    Rabbit monoclonal [EPR16642] to Bcl-XL
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, IHC-P, WB, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment corresponding to Human Bcl-XL aa 1 to the C-terminus.
    Database link: Q07817

  • Positive control
    • WB: Ramos, Jurkat, HeLa or K562 whole cell lysate. Mouse brain, heart, kidney, spleen. Rat brain, heart, kidney, spleen. C6, RAW 264.7, PC-12 or NIH/3T3 whole cell lysate. IHC-P: Human kidney, mouse colon, rat testis. ICC/IF: HeLa cells.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab178844 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
Flow Cyt 1/100.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/20000. Detects a band of approximately 30 kDa (predicted molecular weight: 26 kDa).
ICC/IF 1/1000.
IP 1/70.

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

Images

  • All lanes : Anti-Bcl-XL antibody [EPR16642] (ab178844) at 1/20000 dilution

    Lane 1 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
    Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 4 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Developed using the ECL technique.

    Predicted band size: 26 kDa



    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM /TBST

  • Immunofluorescence analysis of 4% paraformaldehyde fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Bcl-XL/S (green) with ab178844 at a 1/1000 dilution showing cytoplasmic staining. Permeabilisation using 0.1% tritonX-100.

    Secondary ab: Anti-Rabbit Alexa Fluor® 488 (ab150077) at 1/200 dilution. Counter stain is labeling tubulin (red) with ab7291 at 1/500 dilution with secondary antibody Anti-Mouse AlexaFluor® 594 (ab150120) at 1/400 dilution. DAPI stains the nucleus in blue. -ve control 1 is ab178844 at 1/1000 dilution, Anti-Mouse AlexaFluor® 594 (ab150120) at 1/400 dilution. -ve control 2 is ab7291 at 1/500 dilution, Anti-Rabbit Alexa Fluor® 488 (ab150077) at 1/200 dilution.

  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Bcl-XL with ab178844 at 1:2000 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with hematoxylin. 

    Cytoplasmic staining on epithelial cells of tubules and glomeruli of kidney is observed.

    Negative control: Using PBS instead of primary ab, secondary ab as above.

     

  • Flow Cytometry analysis of HeLa cells labelling Bcl-XL with purified ab178844 at a dilution of 1/100 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Flour® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Bcl-XL with ab178844 at a 1/2000 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with hematoxylin.

    Cytoplasm staining on epithelial cells of mouse colon tissue is observed.

    Negative control: Using PBS instead of primary ab, secondary ab as above.

  • Immunohistochemical analysis of paraffin-embedded Rat tesits labeling Bcl-XL with ab178844 at a 1/2000 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with hematoxylin.

    Cytoplasm staining on epithelial cells of testis is observed.

    Negative control: Using PBS instead of primary ab, secondary ab as above.

     

  • Bcl-XL/S was immunoprecipitated from 1mg of Ramos (Human Burkitt's lymphoma cell line) whole cell extract with ab178844 at 1/70 dilution. Western blot was performed of the immunoprecipitate using ab178844 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Left lane: Ramos whole cell extract. Right lane: PBS instead of Ramos whole cell extract.

    Blocking buffer and concentration: 5% NFDM/TBST. Diluting buffer and concentration:

    5% NFDM/TBST.

  • Immunofluorescence analysis of 4% paraformaldehyde fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells is a comparison between ab178844 and a competitor’s leading rabbit polyclonal antibody.
    Labeling of Bcl-XL/S with ab178844 is conducted at 1/1000 dilution, followed by Anti-Rabbit Alexa Fluor® 488 (ab150077) secondary antibody at 1/200 dilution.
    Labeling of Bcl-XL/S with competitor antibody is conducted at 1/50 dilution, followed by Anti-Rabbit Alexa Fluor® 488 (ab150077) secondary antibody at 1/200 dilution.
    DAPI stains the nucleus in blue.
    -ve control 1 is ab178844 at 1/1000 dilution, Anti-Mouse Alexa Fluor® 594 (ab150120) at 1/400 dilution.
    -ve control 2 is ab7291 at 1/500 dilution, Anti-Rabbit Alexa Fluor® 488 (ab150077) at 1/200 dilution.

  • All lanes : Anti-Bcl-XL/S [EPR16642] antibody (ab178844) at 1/20000 dilution, and a competitor's rabbit polyclonal antibody at 1/100 dilution

    Lane 1 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
    Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 4 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
    developed using the ECL technique

    Predicted band size: 26 kDa
    Observed band size: 30 kDa
    why is the actual band size different from the predicted?



    This western blot image is a comparison between ab178844 and a competitor's leading rabbit polyclonal antibody.

    Blocking buffer and concentration: 5% NFDM/TBST

     

     

  • All lanes : Anti-Bcl-XL/S [EPR16642] antibody (ab178844) at 1/20000 dilution, and a competitor's rabbit polyclonal antibody at 1/100 dilution

    Lane 1 : Mouse brain
    Lane 2 : Mouse heart
    Lane 3 : Mouse kidney
    Lane 4 : Mouse spleen
    Lane 5 : Rat brain
    Lane 6 : Rat heart
    Lane 7 : Rat kidney
    Lane 8 : Rat spleen
    Lane 9 : C6 whole cell lysates
    Lane 10 : Raw264.7 whole cell lysates
    Lane 11 : PC-12 whole cell lysates
    Lane 12 : NIH/3T3 whole cell lysates

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated

    Predicted band size: 26 kDa
    Observed band size: 30 kDa why is the actual band size different from the predicted?



    This western blot image is a comparison between ab178844 and a competitor's leading rabbit polyclonal antibody.

    Blocking buffer and concentration: 5% NFDM/TBST

  • This IHC image is a comparison between ab178844 and a competitor's leading rabbit polyclonal antibody in which both antibodies were tested on Human kidney tissue.

    Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Bcl-XL with ab178844 at 1/2000 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with hematoxylin.
    Cytoplasmic staining on epithelial cells of tubules and glomeruli of kidney is observed.
    Negative control: Using PBS instead of primary ab, secondary ab as above.
    Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Bcl-XL with competitor rabbit polyclonal at 1/100 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with hematoxylin.

  • This IHC image is a comparison between ab178844 and competitor's leading rabbit polyclonal in which both antibodies were tested on Mouse colon tissue.

    Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Bcl-XL with ab178844 at a 1/2000 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with hematoxylin.
    Cytoplasm staining on epithelial cells of mouse colon tissue is observed.
    Negative control: Using PBS instead of primary ab, secondary ab as above.
    Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Bcl-XL with competitor rabbit polyclonal antibody at a 1/100 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with hematoxylin.

  • This IHC image is a comparison between ab178844 and a competitor's leading rabbit polyclonal antibody in which both antibodies were tested on Rat testis tissue.
    Immunohistochemical analysis of paraffin-embedded Rat tesits labeling Bcl-XL with ab178844 at a 1/2000 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with hematoxylin.
    Cytoplasm staining on epithelial cells of testis is observed.
    Negative control: Using PBS instead of primary ab, secondary ab as above.
    Immunohistochemical analysis of paraffin-embedded Rat tesits labeling Bcl-XL with competitor rabbit polyclonal antibody at a 1/100 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with hematoxylin.

  • This immunoprecipitation image is a comparison between ab178844 and a competitor's leading rabbit polyclonal antibody.

    Bcl-XL/S was immunoprecipitated from 1mg of Ramos (Human Burkitt's lymphoma cell line) whole cell extract with ab178844 at 1/70 dilution. Western blot was performed of the immunoprecipitate using ab178844 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Left lane: Ramos whole cell extract. Right lane: PBS instead of Ramos whole cell extract.
    Bcl-XL/S was immunoprecipitated from 1mg of Ramos (Human Burkitt's lymphoma cell line) whole cell extract with competitor rabbit polyclonal antibody at 1/12 dilution. Western blot was performed of the immunoprecipitate using competitor antibody at 1/100 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Left lane: Ramos whole cell extract. Right lane: PBS instead of Ramos whole cell extract.
    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.

References

This product has been referenced in:
  • Saalim M  et al. IL-22 in hepatocyte's survival of Pakistani patients with end stage liver disease: an insight into IL 22 mediated hepato-regenerative pathway. Mol Biol Rep 46:1127-1138 (2019). Read more (PubMed: 30603953) »
  • Xu B  et al. Activation of the MAPK signaling pathway induces upregulation of pro-apoptotic proteins in the hippocampi of cold stressed adolescent mice. Neurosci Lett 699:97-102 (2019). Read more (PubMed: 30711527) »
See all 9 Publications for this product

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