Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23463-30] to CXCR5 - BSA and Azide free
  • Suitable for: ICC, IHC-P, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free
    See all CXCR5 primary antibodies

  • Description

    Rabbit monoclonal [EPR23463-30] to CXCR5 - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    We observe only weak staining in human WB.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: Raji, Daudi, Neuro-2a, A20, Mouse spleen, mouse lymph node and rat lymph node, C6 lysates. IHC-P: Human tonsil tissue. Human diffuse large B- lymphoma tissue. Flow Cyt: Human peripheral blood mononuclear and Raji cells. ICC: Daudi cells.

  • General notes

    ab272936 is the carrier-free version of ab254415. 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.

    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

  • Flow Cytometry - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)
    Flow Cytometry - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)

    This data was developed using the same antibody clone in a different buffer formulation (ab254415)

    Flow cytometry overlay histogram showing wild-type Raji (green line) and CXCR5 knockout Raji cells (ab273380) stained with ab254415 (red line). The cells were incubated in 1x PBS containing 10μg/ml human IgG and 10% normal goat serumto block FC receptors and non-specific protein-protein interaction followed by the antibody (ab254415) (1x106 in 100μl at 0.2 μg/ml) for 30 min at 4°C.

    The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150081) was used at 1/2000 for 30 min at 4°C.

    Isotype control antibody was Rabbit IgG (monoclonal) (ab172730) used at the same concentration and conditions as the primary antibody (wild-type Raji - black line; CXCR5 knockout Raji - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

    Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CXCR5 with ab254415 at 1/5000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human tonsil (PMID: 12393412). The section was incubated with ab254415 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

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

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254415).

  • Western blot - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)
    Western blot - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)
    All lanes : Anti-CXCR5 antibody [EPR23463-30] (ab254415) at 1/1000 dilution

    Lane 1 : Wild-type Raji cell lysate
    Lane 2 : CXCR5 knockout Raji cell lysate
    Lane 3 : Daudi cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 60 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 (ab254415).

    Lanes 1 - 4: Merged signal (red and green). Green - ab254415 observed at 60 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab254415 was shown to react with CXCR5 in Raji wild-type cells in Western blot with loss of signal observed in CXCR5 knockout sample. Wild-type and CXCR5 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab254415 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 - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)
    Immunocytochemistry - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)
    This data was developed using the same antibody clone in a different buffer formulation (ab254415). ab254415 staining CXCR5 in Daudi cells (top panel, positive control) and Jurkat cells (bottom panel, negative control). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab254415 at 5μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
  • Flow Cytometry - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)
    Flow Cytometry - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)

    Flow cytometric analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labelling CXCR5 with ab254415 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254415).

  • Flow Cytometry - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)
    Flow Cytometry - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)

    Flow cytometric analysis of human peripheral blood mononuclear cell (PBMC) cells labelling CXCR5 with ab254415 at 1/500 dilution (0.1µg) (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary at a 1/2000 dilution. Cells were stained with rabbit IgG (Left) or ab254415 (Right), then stained with anti-CD19 conjugated to Alexa Fluor® 647.

    Gated on viable cells.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254415).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)

    Immunohistochemical analysis of paraffin-embedded human diffuse large B-cell lymphoma tissue labeling CXCR5 with ab254415 at 1/5000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human diffuse large B-cell lymphoma. The section was incubated with ab254415 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

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

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254415).

  • Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)
    Anti-CXCR5 antibody [EPR23463-30] - BSA and Azide free (ab272936)

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