PA1-41697

antibody from Invitrogen Antibodies

Targeting: KDM1A AOF2, BHC110, KDM1, KIAA0601, LSD1

Provider product page for PA1-41697

Western blot

Other assay

Antibody data

Antibody Data
Antigen structure
References [1]
Comments [0]
Validations
Western blot [1]
Other assay [4]

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Product number: PA1-41697 - Provider product page
Provider: Invitrogen Antibodies
Product name: LSD1 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: Suggested positive control: brain lysate, human brain protein.
Reactivity: Human, Mouse, Rat, Canine, Zebrafish
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

KDM1A protein structure - PA1-41697 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 Role of S-nitrosylation of Kdm1a in tailfin regeneration in adult zebrafish. A MS/MS fragmentation spectrum for the Cys334-containing peptide of Kdm1a. Peptide sequence is shown at the top left of the spectrum, with the annotation of the identified matched amino terminus-containing ions (b ions) in black and the carboxyl terminus-containing ions (y ions) in red. The spectrum confirms the identity of the peptides CPLYEAN and the labeled C as S-nitrosylated cysteine. B Line graph reporting the quantification of Kdm1a S-nitrosylation (normalized by total Kdm1a) and Kdm1a activity during tailfin regeneration. C Western blotting (WB) for S-nitrosylated Kdm1a in uninjured and at 3, 5, and 10 dpa. Samples were previously immunoprecipitated (IP) for Kdm1a. IP with IgG and input were used as controls. D - F WB for Kdm1a, CoRest and NuRD complexes components following IP with Kdm1a antibody in tailfin uninjured or injured at 5 dpa. Dpa days post-amputation. N = 3 biological replicates.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 Effects of kdm1a knockdown in adult zebrafish. A Western blotting (WB) analysis of Kdm1a control and morpholino KD. Dot plot shows semiquantitative analysis of bands. Two-tailed t -test. B Effects of Kdm1a KD on tailfin regeneration. Dashed red line represents the edge of the resection. Scale bar indicates 2 mm. C Line graph showing changes in tailfin regeneration rate following kdm1a KD. Two-way ANOVA followed by Bonferroni's multiple comparisons test. D , E WB for H3K4unme (unmethylated), H3K4me1, H3K4me2, and H3K4me3 in control uninjured, injured and injured + kdm1a KD at 5 dpa. Dot plot shows semi-quantitative analysis of bands. Histone H3 was used as loading control. Dpa days post-amputation. Two-way ANOVA followed by Bonferroni's multiple comparisons test, p values indicate comparisons of uninjured vs. other groups. N = 3 biological replicates. Data are presented as mean values +/- SEM.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 5 Analysis of S-nitrosylation in endothelial cells during tailfin regeneration. A Brightfield and fluorescence images of Tg(fli1:EGFP) y1 zebrafish tailfin at 3 days post-amputation (dpa) showing formation of new vessel branches (GFP signal). Scale bar measures 500 mum. B FACS plot of GFP + and GFP - cells in the tailfin in control and during regeneration were separated by FACS. C Quantification of GFP+ cells as shown in FACS plots. Two-tailed t -test. D Western blotting (WB) of total S-nitrosylated proteins in zebrafish tailfin endothelial (GFP+) cells. E WB of Kdm1a and S-nitrosylated Kdm1a in endothelial (GFP+) cells control, injury and injury + PTIO (NO scavenger) 10 mM. Dot plot shows semi-quantitative analysis. p values vs. 5 dpa group. F Vessel density analysis in Tg(fli1:EGFP) y1 zebrafish tailfin uninjured, injured and injured + PTIO 10 mM, measured as total length of vessels. G ChIP-PCR analysis in GFP + cells isolated from the regenerating tailfin showing H3K4me2-binding complex with vegfaa and tek promoters. Rabbit IgG were used as a negative control. H Real time PCR analysis for endothelial genes in GFP+ cells from zebrafish control, injury and injury + PTIO 10 mM. Histone H3 was used as loading control. One-way ANOVA followed by Bonferroni's multiple comparisons test, p values indicate comparisons vs. uninjured. N = 3 biological replicates. Data are presented as mean values +/- SEM.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 6 Modulation of Kdm1a S-nitrosylation during tailfin regeneration. A Kdm1a mRNA C334A was generated by site-directed mutagenesis, replacing the aa Cys334 with Ala. B - D Zebrafish embryos injected with kdm1a morpholino (Mo), or co-injected with kdm1a Mo with kdm1a mRNA C334A or wild type. B Western blotting and semi-quantitative analysis showed the effective knockdown and rescue of kdm1a following the different treatments. beta-tubulin was used as loading control. C , D Images and dot plot of tailfin regeneration following kdm1a modulation ( p values vs. control). Scale bar measures 100 mum. E Working model. Tissue injury promotes the S-nitrosylation of the Cys334 of Kdm1a. S-nitrosylated Kdm1a detaches from the CoRest complex and loses its demethylase activity on H3K4. One-way ANOVA followed by Bonferroni's multiple comparisons test, p values indicate comparisons vs. control. N = 3 biological replicates. Data are presented as mean values +/- SEM.