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    bcl-xl-antibody-e18-bsa-and-azide-free-ab199099.pdf

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Cell Biology Apoptosis Intracellular Bcl2 Family
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RecombinantRabMAb

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

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  • Certificate of Compliance
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Western blot - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
  • Immunocytochemistry/ Immunofluorescence - 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)
  • Flow Cytometry (Intracellular) - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
  • Immunoprecipitation - 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)
  • Immunocytochemistry/ Immunofluorescence - 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)
  • Flow Cytometry (Intracellular) - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
  • OI-RD Scanning - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
  • Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)

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: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WB
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Unconjugated

You may also be interested in

Protein
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Recombinant Human Bcl-XL protein (His tag) (ab180062)
Primary
Product image
Anti-MCL1 antibody [Y37] - BSA and Azide free (ab186822)
Conjugation
Product image
APC Conjugation Kit - Lightning-Link® (ab201807)

View more associated products

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: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WBmore 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.

  • 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.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    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.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Dissociation constant (KD)

    KD = 6.50 x 10 -11 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E18
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Bcl2 Family
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Bcl 2 family
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Cell survival & death
    • Cancer
    • Tumor biomarkers
    • Oncoproteins
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Bcl 2 family
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism

Associated products

  • Alternative Versions

    • HRP Anti-Bcl-XL antibody [E18] (ab200600)
    • PE Anti-Bcl-XL antibody [E18] (ab208747)
    • Alexa Fluor® 647 Anti-Bcl-XL antibody [E18] (ab303457)
    • Alexa Fluor® 555 Anti-Bcl-XL antibody [E18] (ab306571)
    • Anti-Bcl-XL antibody [E18] (ab32370)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Jurkat whole cell lysate (ab30128)
    • NIH/3T3 whole cell lysate (ab7179)
    • RAW 264.7 whole cell lysate (ab7187)
    • Jurkat whole cell lysate (ab7899)
    • K-562 whole cell lysate (ab7911)
  • Recombinant Protein

    • Recombinant Human Bcl-XL protein (His tag) (ab180062)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
Use at an assay dependent concentration.

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

ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
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.

Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.

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

ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
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.
  • Target information above from: UniProt accession Q07817 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • SwissProt: P53563 Rat
    • Unigene: 516966 Human
    • Unigene: 238213 Mouse
    • Unigene: 10323 Rat
    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 (Intracellular) - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    Flow Cytometry (Intracellular) - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)

    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. 

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

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

     

     

  • OI-RD Scanning - Anti-Bcl-XL antibody [E18] - BSA and Azide free (ab199099)
    OI-RD Scanning - 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).

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

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (17)

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

ab199099 has been referenced in 17 publications.

  • Gao J  et al. Inhibition of esophageal-carcinoma cell proliferation by genistein via suppression of JAK1/2-STAT3 and AKT/MDM2/p53 signaling pathways. Aging (Albany NY) 12:6240-6259 (2020). PubMed: 32276266
  • 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
View all Publications for this product

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