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Antibodies

AbFlex® KDM5A antibody (rAb)

Catalog No: 91211 Format: 100 µg $375 Buy Now
Catalog No: 91212 Format: 10 µg $75 Buy Now

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Antibody Type:Recombinant
Isotype:IgG2a
Purification:Protein A Chromatography
Host:Mouse
Molecular Weight:196 kDa
Reactivity:Human

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated

for AbFlex KDM5A antibody (rAb)Application Notes


Validated Applications:
ChIP-Seq: 4 µg per ChIP
WB: 0.5 - 2 µg/mL

For optimal results, we recommend the addition of 0.1% Tween 20 to all Blotto solutions to reduce background. Individual optimization may be required.

Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western Blot.

for AbFlex KDM5A antibody (rAb)Immunogen

This antibody was raised against full length, recombinant human KDM5A.

for AbFlex KDM5A antibody (rAb)Buffer

Purified IgG in 140 mM Hepes, pH 7.5, 70 mM NaCl, 32 mM NaOAc, 0.035% sodium azide, 30% glycerol.

 
AbFlex<sup>®</sup> KDM5A antibody (rAb) tested by ChIP-Seq.

AbFlex® KDM5A antibody (rAb) tested by ChIP-Seq
Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 25 µg of chromatin from human H727 lung cell chromatin and 4 µg of AbFlex KDM5A antibody. ChIP DNA was sequenced on the Illumina NextSeq and 13 million sequence tags were mapped to identify KDM5A binding sites on chromosome 8. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

Specificity of AbFlex® KDM5A demonstrated by Western blot.

AbFlex® KDM5A antibody (rAb) tested by Western blot.
40 µg of HeLa cell nuclear extract was run on SDS-PAGE and probed with AbFlex® KDM5A antibody at 2 µg/ml.

MW: 196 kDa

Specificity of AbFlex® KDM5A demonstrated by Western blot.

Specificity of AbFlex® KDM5A demonstrated by Western blot.
500 ng of various FLAG-tagged human full-length recombinant proteins was loaded per lane and probed with KDM5A antibody at 1µg/ml (top panel) or FLAG antibody (bottom panel). KDM5A antibody only detects KDM5A protein; the FLAG antibody shows the presence of all proteins on the blot.
Lane 1: KDM3A (Cat. No. 31456). Lane 2: KDM3B (Cat. No. 31429). Lane 3: KDM4A (Cat. No. 31457). Lane 4: KDM4B Cat. No. 31501). Lane 5: KDM4C (Cat. No. 31458). Lane 6: KDM4D (Cat. No. 31459). Lane 7: KDM5A (Cat. No. 31431). Lane 8: KDM5B (Cat. No. 31432). Lane 9: KDM5C (Cat. No. 31433).

for AbFlex KDM5A antibody (rAb)Background

AbFlex® antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6XHis tag, an avidin tag sequence for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports.  KDM5A Lysine (K)-specific demethylase 5A (KDM5A), also known as Jumonji, AT rich interactive domain 1A (JARID1A), is a histone demethylase that specifically demethylates lysine 4 of histone H3 (H3K4), thereby playing a central role in defining the histone code. KDM5A demethylates trimethylated and dimethylated, but not monomethylated H3K4. It does not demethylate histone H3K9, H3K27, H3K36, H3K79or H4K20. KDM5A may function to stimulate transcription mediated by nuclear receptors and may be involved in transcriptional regulation of HOX proteins during cell differentiation. Also, KDM5A may be involved in the transcriptional repression of cytokines such as CXCL12. It has been shown to bind directly with the Retinoblastoma (pRb) protein which regulates cell proliferation. KDM5A also interacts with Rhombotin-2 which functions distinctly in erythropoiesis and in T-cell leukemogenesis.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with AbFlex® KDM5A antibody (rAb).

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use AbFlex® KDM5A antibody (rAb).

for AbFlex KDM5A antibody (rAb)Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

for AbFlex KDM5A antibody (rAb)Guarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting
 

Technical Data Sheet

AbFlex® KDM5A antibody (rAb)
 

Data Thumbnails

AbFlex<sup>®</sup> KDM5A antibody (rAb) tested by ChIP-Seq.

ChIP-Seq of AbFlex® KDM5A antibody (rAb).
(Click image to enlarge and see details.) You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

Specificity of AbFlex® KDM5A demonstrated by Western blot.

Specificity of AbFlex® KDM5A demonstrated by Western blot.
(Click to enlarge and view details)

Specificity of AbFlex® KDM5A demonstrated by Western blot.

Specificity of AbFlex® KDM5A demonstrated by Western blot.
(Click to enlarge and view details)