Anti-Histone H1.2 Rabbit mAb - ChIP, CUT&RUN and CUT&Tag Grade

Product

Anti-Histone H1.2 Rabbit mAb - ChIP, CUT&RUN and CUT&Tag Grade

Catalog number :AT0728

Histones are basic nuclear proteins responsible for nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene is intronless and encodes a member of the histone H1 family. Transcripts from this gene lack polyA tails but instead contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6. [provided by RefSeq]

UniProt ID:

P16403

Overview

Reactivity: Human, mouse, rat, bovine, chicken, Drosophilia melanogaster, Zebrafish

Tested applications: Western Blotting 1:1000

Immunofluorescence 1:500

Immunohistochemistry (Paraffin) 1:500

Immunoprecipitation(IP) 1;100
ChIP, CUT&RUN and CUT&Tag

Optimal dilutions/concentrations should be determined by the end user.

Specificity: This antibody recognizes endogenous levels of total histone H1.2 protein. This antibody does not cross-react with other histone proteins.

Properties

Immunogen: Recombinant protein encompassing a sequence within the center region of human Histone H1.2. The exact sequence is proprietary.

Clonality: Monoclonal, clone number: 4GC21

Isotype: Rabbit IgG

Form: Liquid, 100 μl,1mg/ml, PBS (pH 7.2) and 40% Glycerol，0.02% Sodium Azide

Storage instruction: Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C, Avoid freeze / thaw cycle.

Host: Rabbit

Applications

WB Image: All lanes: Anti-Histone H1.2 antibody at 1/2000 dilution

Lane 1: HeLa cell lysate

Lane 2: 293 cell lysate

Lane 3: A375 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary

All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 21 kDa

Observed band size: 29 kDa

Why is the actual band size different from the predicted?

Western blotting is a technique that separates proteins based on size - in general, the smaller the protein the faster it migrates through the gel. However, migration is also affected by other factors and so the actual band size observed may differ from that predicted. Common factors include:

1, post-translational modification - e.g. phosphorylation, glycosylation etc which increases the size of the protein

2, post-translation cleavage - e.g. many proteins are synthesized as pro-proteins, and then cleaved to give the active form, e.g. pro-caspases

3, splice variants - alternative splicing may create different sized proteins from the same gene

4, relative charge - the composition of amino acids (charged vs non-charged)

5, multimers - e.g. dimerisation of a protein. This is usually prevented in reducing conditions, although strong interactions can result in the appearance of higher bands

IHC Image: Immunohistochemistry-Paraffin: Histone H1.2 Antibody - Immunohistochemical analysis of paraffin-embedded Colon ca, using antibody at 1:500 dilution.

ICC/IF Image: Immunocytochemistry/Immunofluorescence: Histone H1.2 Antibody- Confocal immunofluorescence analysis of paraformaldehyde-fixed HeLa, using Histone H1. 2 antibody (Green) at 1:500 dilution. Alpha-tubulin filaments are labeled with Alpha-tubulin antibody (Red) at 1:2000.

Anti-Histone H1.2 Rabbit mAb - ChIP, CUT&RUN and CUT&Tag Grade

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