Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-TNF alpha antibody [EPR19147] - BSA and Azide free (ab271939)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19147] to TNF alpha - BSA and Azide free
  • Suitable for: ELISA, WB, IP, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-TNF alpha antibody [EPR19147] - BSA and Azide free
    See all TNF alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR19147] to TNF alpha - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ELISA, WB, IP, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: TPA pretreated THP-1 treated with 100 ng/ml LPS for 7 hours and TPA pretreated THP-1 treated with 100 ng/ml LPS for 4 hours, then added 1 µg/ml BFA for 3 hours whole cell lysates; TPA pretreated RAW 264.7, TPA pretreated RAW 264.7 treated with 100 ng/ml LPS for 7 hours and TPA pretreated RAW 264.7 treated with 100 ng/ml LPS for 4 hours, then added 1 µg/ml BFA for 3 hours whole cell lysates. IP: TPA pretreated THP-1 treated with 100 ng/ml LPS for 4 hours, then added 1 µg/ml BFA for 3 hours cell lysate.

  • General notes

    ab271939 is the carrier-free version of ab183218. 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

    This product 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.

Properties

Images

  • Western blot - Anti-TNF alpha antibody [EPR19147] - BSA and Azide free (ab271939)
    Western blot - Anti-TNF alpha antibody [EPR19147] - BSA and Azide free (ab271939)
    All lanes : Anti-TNF alpha antibody [EPR19147] (ab183218) at 1/1000 dilution

    Lane 1 : Wild-type THP-1 Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 2 : Wild-type THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 3 : TNF alpha knockout THP-1 Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 4 : TNF alpha knockout THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 5 : U937 PMA treated (10 mM, 2 days) plus 16 h no treatment and Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 6 : U937 PMA treated (10 mM, 2 days) and LPS treated (1 µg/ml, 16 h) plus Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 25 kDa
    Observed band size: 26 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab183218).

    Lanes 1 - 6: Merged signal (red and green). Green - ab183218 observed at 26 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab183218 was shown to react with TNF alpha in THP-1 wild-type cells in Western blot with loss of signal observed in TNF knockout sample. Wild-type and TNF knockout THP-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab183218 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [EPR19147] - BSA and Azide free (ab271939)
    Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [EPR19147] - BSA and Azide free (ab271939)

    Immunocytochemistry/ Immunofluorescence analysis of RAW264.7(Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml LPS for 7 h and 1µg/ml BFA for the last 3h cells labeling TNF alpha with purified ab183218 at 1/5000 dilution (0.09 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183218).
  • ELISA - Anti-TNF alpha antibody [EPR19147] - BSA and Azide free (ab271939)
    ELISA - Anti-TNF alpha antibody [EPR19147] - BSA and Azide free (ab271939)

    ELISA using ab183218 between 0.1 and 1000 ng/ml to detect TNF alpha peptide. Secondary used was Alkaline Phosphatase AffiniPure Goat Anti-Rabbit IgG (H+L) 1/2000.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183218).

  • Immunoprecipitation - Anti-TNF alpha antibody [EPR19147] - BSA and Azide free (ab271939)
    Immunoprecipitation - Anti-TNF alpha antibody [EPR19147] - BSA and Azide free (ab271939)

    TNF alpha was immunoprecipitated from 0.35 mg of TPA pretreated THP-1 (Human monocytic leukemia cell line) whole cell lysate treated with 100 ng/ml LPS for 4 hours, then added 1 μg/ml BFA for 3 hours with ab183218 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab183218 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: TPA pretreated THP-1 whole cell lysate treated with 100 ng/ml LPS for 4 hours, then added 1 μg/ml BFA for 3 hours 10µg (Input).

    Lane 2: ab183218 IP in TPA pretreated THP-1 whole cell lysate treated with 100 ng/ml LPS for 4 hours, then added 1 μg/ml BFA for 3 hours.

    Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab183218 in TPA pretreated THP-1 whole cell lysate treated with 100 ng/ml LPS for 4 hours, then added 1 μg/ml BFA for 3 hours.

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

    Exposure time: 1 second.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183218).
  • Anti-TNF alpha antibody [EPR19147] - BSA and Azide free (ab271939)
    Anti-TNF alpha antibody [EPR19147] - BSA and Azide free (ab271939)

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