- 720035 - Invitrogen Antibodies JUND antibody

Submitted references IL-2/IL-7-inducible factors pioneer the path to T cell differentiation in advance of lineage-defining factors.

Bevington SL, Keane P, Soley JK, Tauch S, Gajdasik DW, Fiancette R, Matei-Rascu V, Willis CM, Withers DR, Cockerill PN
The EMBO journal 2020 Nov 16;39(22):e105220

TPC2-mediated Ca(2+) signaling is required for axon extension in caudal primary motor neurons in zebrafish embryos.

Guo C, Webb SE, Chan CM, Miller AL
Journal of cell science 2020 Jul 10;133(13)

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (20 µg lysate) of Cos7 (Lane 1), K562 (Lane 2), A431 (Lane 3) and Jurkat (Lane 4). The blots were probed with Anti-JunD Recombinant Rabbit Polyclonal Antibody (Product # 720035, 0.5-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 40 kDa band corresponding to JunD was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (20 µg lysate) of Cos7 (Lane 1), K562 (Lane 2), A431 (Lane 3) and Jurkat (Lane 4). The blots were probed with Anti-JunD Recombinant Rabbit Polyclonal Antibody (Product # 720035, 0.5-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 40 kDa band corresponding to JunD was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence was performed on fixed and permeabilized A-431cells for detection of JunD using Anti-JunD Rabbit Polyclonal Antibody (Product # 720035, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 0.4 µg/mL, 1:2500). Panel a) shows representative cells that were stained for detection and localization of JunD protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938, 1:50). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 594 Phalloidin (Product # A12381, 1:200). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of JunD. Panel e) represents control cells with no primary antibody to assess background.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow Cytometry analysis of JunD was performed on HeLa cells labeled with Anti-JunD Rabbit Polyclonal Antibody (Product# 720035, 2-4 ug/ 1M cells) or with rabbit isotype control and detected with Goat anti-Rabbit IgG (H+L) Superclonalª Secondary Antibody, Alexa Fluor¨ 488 conjugate (Product # A27034, 0.4 ug/ml, 1:2500) as represented by the red and pink histograms respectively. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control. A representative 10,000 cells were acquired and analyzed for each sample using an Attune¨ Acoustic Focusing Cytometer (4468770).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Chromatin Immunoprecipitation (ChIP) was performed using Anti-JunD Rabbit Polyclonal Antibody (Product # 720035, 5 µg) on sheared chromatin from 2 million HeLa cells (serum starved for 24 hours and serum released for 2 hours) using the "MAGnify ChIP system" kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by 7500 Fast qPCR system (Product # 4351106) with optimized PCR primer pairs for the promoters of the active VEGF, CREB5 genes, used as positive control target gene, and the promoter region of the GAPDH, used as negative control target gene. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 3 Figure IL-2 pDHSs bind IL-2 regulated factors De novo identification of DNA consensus motifs enriched in the 1,000 IL-2 pDHSs determined by HOMER. Jund mRNA levels in T B IL-2 and T B IL-2nil. Standard deviation is shown from the mean of 3 samples. P -values were calculated from the RNA-Seq data using Limma with Benjamini-Hochberg correction for multiple testing. Western blot analyses showing JUND and pSTAT5 protein levels in T B IL-2 and T B IL-2nil (left panel) with the signal calculated from 2 replicates relative to T B IL-2 in the right panel. DNA motif density plots (right) showing the location of binding motifs for RUNX, ETS, STAT, and AP-1 transcription factors at the 32,694 DHSs detected in T B IL-2 and T B IL-2nil, as depicted in Fig 2A, ordered according to fold enrichment of DNase-Seq tag counts in T B IL-2 compared to T B IL-2nil (left). DNA sequence tag density plots from ChIP-Seq assays of RUNX1, ETS1, STAT5, and JUND in T B IL-2, and STAT5 and JUND in T B IL-2nil ordered as in (D). UCSC genome browser tracks at the Il13/Il4/Il5 Th2 cytokine locus LCR (indicated by the gray box within the Rad50 gene), Gzmb and Il24 showing DNase-Seq and ChIP-Seq for STAT5 and JUND in T B IL-2 and T B IL-2nil. Replicate JUND and STAT5 ChIP-Seq tracks are shown for T B IL-2.

720035

Antibody data