Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PU.1/Spi1 antibody [EPR22624-20] - ChIP Grade – BSA and Azide free (ab264473)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22624-20] to PU.1/Spi1 - ChIP Grade – BSA and Azide free
  • Suitable for: ICC/IF, WB, IHC-P, Flow Cyt, IP, ChIP
  • Reacts with: Mouse

Overview

  • Product name

    Anti-PU.1/Spi1 antibody [EPR22624-20] - ChIP Grade – BSA and Azide free
    See all PU.1/Spi1 primary antibodies

  • Description

    Rabbit monoclonal [EPR22624-20] to PU.1/Spi1 - ChIP Grade – BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Mouse
    Flow Cyt
    Mouse
    ICC/IF
    Mouse
    IHC-P
    Mouse
    IP
    Mouse
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: RAW264.7, NIH/3T3 and J774A.1 lysates. IHC-P: Mouse spleen and Mouse liver tissues. ICC/IF: RAW 264.7 cells. Flow Cyt: RAW264.7 cells. IP: J774A.1 cells.

  • General notes

    ab264473 is the carrier-free version of ab227835. 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

    ab264473 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • ChIP - Anti-PU.1/Spi1 antibody [EPR22624-20] - BSA and Azide free (ab264473)
    ChIP - Anti-PU.1/Spi1 antibody [EPR22624-20] - BSA and Azide free (ab264473)

    Chromatin was prepared from J774A.1 cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30 mins and then formaldehyde for 10 min. 
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab227835 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).

    Primers and probes are from paper PMID: 11869689
    *https://www.abcam.com/resources?keywords=X%20ChIP%20protocol

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227835).

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PU.1/Spi1 antibody [EPR22624-20] - BSA and Azide free (ab264473)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PU.1/Spi1 antibody [EPR22624-20] - BSA and Azide free (ab264473)

    Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Spi1 with ab227835 at 1/5000 dilution (0.103 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in Kupffer cells of moue liver. The section was incubated with ab227835 for 15 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control/ Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227835).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PU.1/Spi1 antibody [EPR22624-20] - BSA and Azide free (ab264473)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PU.1/Spi1 antibody [EPR22624-20] - BSA and Azide free (ab264473)

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Spi1 with ab227835 at 1/5000 dilution (0.103 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse spleen. The section was incubated with ab227835 for 15 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control/ Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227835).

  • Immunoprecipitation - Anti-PU.1/Spi1 antibody [EPR22624-20] - BSA and Azide free (ab264473)
    Immunoprecipitation - Anti-PU.1/Spi1 antibody [EPR22624-20] - BSA and Azide free (ab264473)

    Spi1 was immunoprecipitated from 0.35 mg J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate with ab227835 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab227835 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

    Lane 1: J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate 10ug

    Lane 2: ab227835 IP in J774A.1 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab227835 in J774A.1 whole cell lysate

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

    Exposure time: 30 seconds

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227835).

  • Flow Cytometry - Anti-PU.1/Spi1 antibody [EPR22624-20] - BSA and Azide free (ab264473)
    Flow Cytometry - Anti-PU.1/Spi1 antibody [EPR22624-20] - BSA and Azide free (ab264473)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling Spi1 with ab227835 at 1/50 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution 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 (ab227835).

  • Immunocytochemistry/ Immunofluorescence - Anti-PU.1/Spi1 antibody [EPR22624-20] - BSA and Azide free (ab264473)
    Immunocytochemistry/ Immunofluorescence - Anti-PU.1/Spi1 antibody [EPR22624-20] - BSA and Azide free (ab264473)

    Immunofluorescent analysis of 4% parafoprmal;dehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling PU.1/Spi1 with ab227835 at 1/500 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). Confocal image showing nuclear staining in RAW 264.7 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).

    Secodary antibody only control: Used PBS instead of primary antibody, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227835).

  • Anti-PU.1/Spi1 antibody [EPR22624-20] - ChIP Grade – BSA and Azide free (ab264473)
    Anti-PU.1/Spi1 antibody [EPR22624-20] - ChIP Grade – BSA and Azide free (ab264473)

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