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ab38771 |
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Anti-PADI4 / PAD4 antibody
See all PADI4 / PAD4 products (8) ...
Rabbit polyclonal to PADI4 / PAD4
IHC-P, ICC/IF, WBmore details
Reacts with
Human
Does not react with
Mouse
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human PADI4 / PAD4.
(Peptide available as ab38771.)
This antibody gave a positive result in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) A431 (Human epithelial carcinoma cell line) HEK 293 (Human embryonic kidney cell line) HepG2 (Human hepatocellular liver carcinoma cell line)
Liquid
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
Immunogen affinity purified
Polyclonal
IgG
Our Abpromise guarantee covers the use of ab38772 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-P: Use at an assay dependent concentration.
ICC/IF: Use a concentration of 1 µg/ml
WB: 1/250Detects a band of approximately 67 kDa (predicted molecular weight: 74 kDa).
In vivo expression of PADI4 correlates with the appearance of citrulline in histones. It has been demonstrated that PADI4 converts arginines in the histone H3 tail to citrulline, blocks transcriptional activation and is recruited to the endogenous promotor. PADI4 represents a novel class of histone modifying enzymes probably linked to oestrogen regulated transcription. PADI4 is thought to be a rheumatoid arthritis susceptibility locus, and could have an important role in the pathogenesis of rheumatoid arthritis by increasing citrullination of proteins in rheumatoid arthritis synovial tissues, leading, in a cytokine-rich milieu, to a break in tolerance to citrullinated peptides processed and presented in the appropriate HLA context.
Cytoplasm. Nucleus. Cytoplasmic granule. Note: Cytoplasmic granules of eosinophils and neutrophils.
Western blot - PADI4 / PAD4 antibody (ab38772)

All lanes : Anti-PADI4 / PAD4 antibody (ab38772) at 1/250 dilution
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)
Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate (ab7902)
Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate (ab7900)
Lysates/proteins at 10 µg per lane.
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 74 kDa
Observed band size : 67 kDa (why is the actual band size different from the predicted?)
Full length PADI4 has been shown to migrate at 67 kDa. Please refer to the reference below for further details.
Chang X et al. Localization of peptidylarginine deiminase 4 (PADI4) and citrullinated protein in synovial tissue of rheumatoid arthritis. Rheumatology (Oxford) 44:40-50 (2005). PMID:15466895.
Immunocytochemistry/ Immunofluorescence - PADI4 / PAD4 antibody (ab38772)

ICC/IF image of ab38772 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab38772, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PADI4 / PAD4 antibody (ab38772)

ab38772 staining PADI4 / PAD4 in human spleen tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with 10% NBF and a heat mediated antigen retrieval step was performed using TRIS/EDTA pH 9.0. Samples were then blocked with 0.25% casein in PBS for 5 minutes at 25°C followed by incubation with the primary antibody at a 1/400 dilution for 30 minutes at 25°C (serum free antibody diluent). An undiluted HRP-conjugated goat polyclonal was used as secondary antibody.
Image courtesy of Ross Benik by Abreview.
This product has been referenced in:
See 1 publication for this product
Publishing research using ab38772? Please let us know so that we can cite the reference in this datasheet
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All lanes : Anti-PADI4 / PAD4 antibody (ab38772) at 1/250 dilution
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)
Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate (ab7902)
Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate (ab7900)
Lysates/proteins at 10 µg per lane.
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 74 kDa
Observed band size : 67 kDa (why is the actual band size different from the predicted?)
Full length PADI4 has been shown to migrate at 67 kDa. Please refer to the reference below for further details.
Chang X et al. Localization of peptidylarginine deiminase 4 (PADI4) and citrullinated protein in synovial tissue of rheumatoid arthritis. Rheumatology (Oxford) 44:40-50 (2005). PMID:15466895.

ICC/IF image of ab38772 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab38772, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

ab38772 staining PADI4 / PAD4 in human spleen tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with 10% NBF and a heat mediated antigen retrieval step was performed using TRIS/EDTA pH 9.0. Samples were then blocked with 0.25% casein in PBS for 5 minutes at 25°C followed by incubation with the primary antibody at a 1/400 dilution for 30 minutes at 25°C (serum free antibody diluent). An undiluted HRP-conjugated goat polyclonal was used as secondary antibody.
Image courtesy of Ross Benik by Abreview.

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