PA5-46262

antibody from Invitrogen Antibodies

Targeting: SPRTN C1orf124, DKFZP547N043, DVC1, Spartan

Provider product page for PA5-46262

Western blot

Other assay

Antibody data

Antibody Data
Antigen structure
References [1]
Comments [0]
Validations
Western blot [1]
Other assay [2]

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Product number: PA5-46262 - Provider product page
Provider: Invitrogen Antibodies
Product name: SPRTN Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: Peptide sequence: QIKSSGNDPK YSTTTAQNSS SSSSQSKMVN CPVCQNEVLE SQINEHLDWC Sequence homology: Human: 100%
Reactivity: Human
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 0.5 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

SPRTN protein structure - PA5-46262 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 USP11 is required for cell survival upon DNA damage by DPC-inducing agents. A , WB showing knockdown efficiency of USP11 in U2OS cells stably expressing nonsilencing shRNA control or two different shRNA sequences (sh1 and sh2) targeted to USP11. B - F , clonogenic cell survival curves for shRNA control and USP11 knockdown U2OS treated with indicated concentrations of ( B ) CPT for 24 h, ( C ) VP16 for 4 h, ( D ) formaldehyde for 20 min, ( E ) HU for 24 h, or ( F ) MMC for 24 h. G , WB showing knockdown efficiency of USP11 and SPRTN. H and I , clonogenic cell survival curves for USP11 and SPRTN single and double -knockdown cells treated with indicated concentrations of ( H ) CPT for 24 h or ( I ) VP16 for 2 h. B - F , H and I , percent cell survival was calculated. Data are presented as mean +- SD (n = 3). Statistical analysis: Two-tailed paired t -test was performed using confidence interval =90% and alpha = 0.1; * p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 USP11 deubiquitinates SPRTN upon DPC induction. A and B , HEK 293T ( A ) or HCT116 ( B ) cells stably expressing shRNA targeted to USP11 were transfected with SFB-SPRTN. Twenty-four hours later, cells were treated with the indicated concentrations of formaldehyde for 2 h. C , HCT116 cells stably expressing shRNA targeted to USP11 were treated with the indicated concentrations of formaldehyde for 1 h. The nuclear fraction was isolated and used for WB. D , A549 cells were transfected with siRNA targeted to USP11. Seventy-two hours later, cells were treated with the indicated concentrations of formaldehyde for 2 h. A - D , Top panel , cell lysates were immunoblotted with the indicated antibodies. Bottom panel , graph shows percent total FLAG-SPRTN or percent total SPRTN quantified from anti-FLAG or anti-SPRTN blots, respectively. The percent of unmodified and monoubiquitinated SPRTN was calculated. E , HEK 293T cells were transfected with SFB-SPRTN alone or in combination with Myc-USP11 FL or C318S mutant. Left panel , cell lysates were immunoprecipitated with S-protein agarose beads, and immunoblotting was performed using the indicated antibodies. Anti-Ub blot was probed with K63 ubiquitin antibody. Right panel , graph showing quantification of SFB-SPRTN auto-cleavage products. Anti-FLAG high exposure blot was used for quantification. WB, western blot. See also Figs. S4 , S5 , and S6 .