Recombinant
RabMAb

Recombinant Anti-AICDA antibody [EPR23436-45] - ChIP Grade – BSA and Azide free (ab269457)

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Overview

  • Product name

    Anti-AICDA antibody [EPR23436-45] - ChIP Grade – BSA and Azide free
    See all AICDA primary antibodies

  • Description

    Rabbit monoclonal [EPR23436-45] to AICDA - ChIP Grade – BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP, ChIPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human AICDA aa 150 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9GZX7

  • Positive control

    • WB: Ramos,NAMALWA and Raji lysates. IHC-P: Human tonsil, Human Hodgkin lymphoma and Human diffuse large B-cell lymphoma tissues. ICC/IF: NAMALWA cells. FC: NAMALWA cells. IP: Ramos and NAMALWA cells. ChIP: Chromatin prepared from Ramos cells.

  • General notes

    ab269457 is the carrier-free version of ab269454. 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 <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab269457 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

Applications

Our Abpromise guarantee covers the use of ab269457 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
WB Use at an assay dependent concentration. Detects a band of approximately 24 kDa (predicted molecular weight: 23 kDa).
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.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration.

Target

  • Function

    RNA-editing deaminase involved in somatic hypermutation, gene conversion, and class-switch recombination. Required for several crucial steps of B-cell terminal differentiation necessary for efficient antibody responses.

  • Tissue specificity

    Strongly expressed in lymph nodes and tonsils.

  • Involvement in disease

    Defects in AICDA are the cause of hyper-IgM immunodeficiency syndrome type 2 (HIGM2) [MIM:605258]; also known as hyper-IgM syndrome 2. HIGM2 is an autosomal recessive disorder characterized by normal or elevated serum IgM levels with absence of IgG, IgA, and IgE, resulting in a profound susceptibility to bacterial infections. HIGM2 causes the absence of Ig class switch recombination (CSR), the lack of Ig somatic hypermutations, and lymph node hyperplasia caused by the presence of giant germinal centers.

  • Sequence similarities

    Belongs to the cytidine and deoxycytidylate deaminase family.
  • Information by UniProt

  • Database links

    • Alternative names

      • Activation induced cytidine deaminase antibody
      • Activation induced deaminase antibody
      • Activation-induced cytidine deaminase antibody
      • AICDA antibody
      • AICDA_HUMAN antibody
      • AID antibody
      • ARP 2 antibody
      • ARP2 antibody
      • CDA 2 antibody
      • CDA2 antibody
      • Cytidine aminohydrolase antibody
      • HIGM2 antibody
      • Integrated into Burkitt's lymphoma cell line Ramos antibody
      see all

    Images

    • ChIP - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)
      ChIP - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)

      Chromatin was prepared from Ramos cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.

      The ChIP was performed with 25 µg of chromatin, 5 µg of ab269454 (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 commercial primers from paper: PMC2905439

      *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 (ab269454).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)

      Immunohistochemical analysis of paraffin-embedded Human diffuse large B-cell lymphoma tissue labeling AICDA with ab269454 at 1/4000 dilution (0.12ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Mainly cytoplasmic staining (weak nuclear staining) in part of tumor cells of human diffuse large B-cell lymphoma (PMID: 29251015).

      The section was incubated with ab255611 for 30 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 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 (ab269454).

    • Immunocytochemistry/ Immunofluorescence - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)
      Immunocytochemistry/ Immunofluorescence - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NAMALWA (human Burkitt's lymphoma B lymphocyte) and K-562 (human chronic myelogenous leukemia lymphoblast) cells labelling AICDA with ab269454 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in NAMALWA cell line. Negative control: K-562 cell line (PMID: 27217538). ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is Ab269454 anti-AICDA 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 (ab269454).

    • Flow Cytometry - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)
      Flow Cytometry - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)

      Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Ramos (human Burkitt's lymphoma B lymphocyte) cells labelling AICDA with ab269454 at 1/500 dilution (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). 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 (ab269454).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)

      Immunohistochemical analysis of paraffin-embedded Human Hodgkin lymphoma tissue labeling AICDA with ab269454 at 1/4000 dilution (0.12ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Mainly cytoplasmic staining (weak nuclear staining) in part of tumor cells of human Hodgkin lymphoma (PMID: 15732141).

      The section was incubated with ab255611 for 30 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 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 (ab269454).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)

      Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling AICDA with ab269454 at 1/4000 dilution (0.12ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Mainly cytoplasmic staining (weak nuclear staining) in germinal center cells of human tonsil (PMID:23877718, 15732141, PMID: 29251015).

      The section was incubated with ab255611 for 30 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 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 (ab269454).

    • Immunoprecipitation - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)
      Immunoprecipitation - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)

      AICDA was immunoprecipitated from 0.35 mg NAMALWA (human Burkitt's lymphoma B lymphocyte) whole cell lysate with ab269454 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab269454 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: NAMALWA whole cell lysate 10ug.

      Lane 2: ab269454 IP in NAMALWA whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab269454 in NAMALWA whole cell lysate.

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

      Exposure time: 90 seconds.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269454).

    • Immunoprecipitation - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)
      Immunoprecipitation - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)

      AICDA was immunoprecipitated from 0.35 mg Ramos (human Burkitt's lymphoma B lymphocyte) whole cell lysate with ab269454 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab269454 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: Ramos whole cell lysate 10ug.

      Lane 2: ab269454 IP in Ramos whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab269454 in Ramos whole cell lysate.

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

      Exposure time: 90 seconds.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269454).

    • Flow Cytometry - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)
      Flow Cytometry - Anti-AICDA antibody [EPR23436-45] - BSA and Azide free (ab269457)

      Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized K-562 (human chronic myelogenous leukemia lymphoblast, Left) / NAMALWA (human Burkitt's lymphoma B lymphocyte, Right) cells labelling AICDA with ab269454 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Blac) isotype control 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.

      Negative control: K-562 cell line (PMID: 27217538).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269454).

    References

    ab269457 has not yet been referenced specifically in any publications.

    Publishing research using ab269457? Please let us know so that we can cite the reference in this datasheet.

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