AbFlex® KDM5A antibody (rAb)

RRID: AB_2793805
Aliases: JARID1A, RBP2; RBBP2; RBBP-2
商品コード: 91211 Format: 100 µg ¥72,000 Buy
商品コード: 91212 Format: 10 µg ¥20,000 Buy

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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

Application Notes

Applications Validated by Active Motif:
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 blocking 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.

Immunogen

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

Buffer

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

 
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 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.

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).

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, a Biotinylation Tag 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, H3K79 or 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).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • 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;
  • NEU = Neutralizing;
  • WB = Western Blotting