Histone H3 (Crotonyl K18 + K23) Recombinant Rabbit mAb (S-R410)

Catalog Number: S0B0853 Application: WB, ICC, ChIP Reactivity: Hu, Ms Conjugation: Unconjugated Brand: Starter

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$100 USD

Size: 25μl 100μl

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Product Details

Product Specification

Host	Rabbit
Antigen	Histone H3 (Crotonyl K18 + K23)
Location	Nucleus
Accession	P68431
Clone Number	S-R410
Antibody Type	Recombinant mAb
Isotype	IgG
Application	WB, ICC, ChIP
Reactivity	Hu, Ms
Purification	Protein A
Concentration	0.5 mg/ml
Conjugation	Unconjugated
Physical Appearance	Liquid
Storage Buffer	PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300
Stability & Storage	12 months from date of receipt / reconstitution, -20 °C as supplied

Dilution

application	dilution	species
WB	1:500	Hu, Ms
ICC	1:500	Hu, Ms
ChIP	1:20-1:50	Hu

Background

Histone H3 (Crotonyl K18 + K23)" refers to Histone H3 that has undergone a particular type of PTM known as crotonylation at two specific lysine residues: lysine 18 (K18) and lysine 23 (K23). Crotonylation is a type of acylation, where an acyl group (in this case, a crotonyl group, derived from crotonic acid) is added to a lysine residue on the histone protein. This modification is thought to have regulatory functions similar to other acylations, such as acetylation, which are often associated with active gene transcription. Crotonylation of histone H3 at K18 and K23 may alter the interaction of histones with DNA and other chromatin-associated proteins, thereby influencing chromatin structure and accessibility to transcription factors and other regulatory complexes.

Picture

Western Blot

WB result of Histone H3 (Crotonyl K18 + K23) Recombinant Rabbit mAb
Primary antibody: Histone H3 (Crotonyl K18 + K23) Recombinant Rabbit mAb at 1/500 dilution
Lane 1: untreated HeLa whole cell lysate 20 µg
Lane 2: HeLa treated with 500 ng/ml TSA for 4 hours whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa

WB result of Histone H3 (Crotonyl K18 + K23) Recombinant Rabbit mAb
Primary antibody: Histone H3 (Crotonyl K18 + K23) Recombinant Rabbit mAb at 1/500 dilution
Lane 1: untreated HeLa whole cell lysate 20 µg
Lane 2: HeLa treated with 30 mM Sodium Butyrate for 4 hours whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa

WB result of Histone H3 (Crotonyl K18 + K23) Recombinant Rabbit mAb
Primary antibody: Histone H3 (Crotonyl K18 + K23) Recombinant Rabbit mAb at 1/500 dilution
Lane 1: NIH/3T3 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa

Immunocytochemistry

ICC shows positive staining in HeLa cells. Anti- Histone H3 (Crotonyl K18 + K23) antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).

ICC shows positive staining in NIH/3T3 cells. Anti- Histone H3 (Crotonyl K18 + K23) antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).

ChIP

Chromatin immunoprecipitation (ChIP) was performed on HeLa + TSA (500ng/ml,4h) cells cross - linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication. Parallel reactions used Histone H3 (Crotonyl K18 + K23) Recombinant Rabbit mAb (S-R410) and Rabbit mAb IgG Isotype Control (SDT-R173) at 1:50 for immunoprecipitation.
Post - immunoprecipitation, both samples were washed, eluted, and cross - links reversed. Purified DNA was analyzed by qPCR.
qPCR (%input: immunoprecipitated DNA/input DNA) showed the enrichment
of RPL30, GAPDH, MYOD1, AFM, SAT-α and SAT-2 in Histone H3 (Crotonyl
K18 + K23) Recombinant Rabbit mAb (S-R410)-immunoprecipitated sample.