PA5-31619

antibody from Invitrogen Antibodies

Targeting: ARHGAP36 FLJ30058

Provider product page for PA5-31619

Western blot

Immunohistochemistry

Other assay

Antibody data

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

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Product number: PA5-31619 - Provider product page
Provider: Invitrogen Antibodies
Product name: ARHGAP36 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant protein fragment
Description: Predicted reactivity: Mouse (91%), Sheep (94%), Bovine (93%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
Reactivity: Human
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 1 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

ARHGAP36 protein structure - PA5-31619 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 ARHGAP36 induces PKAC ubiquitylation and engagement with the ESCRT pathway. ( a ) HEK293T cells were transfected with PKAC-YFP or YFP-Cherry control, Flag-ARHGAP36 and His-Ubiquitin as indicated. Lysates were subjected to GFP IP. ( b ) SILAC-labelled HEK293T cells were transfected with PKAC-Flag in the presence or absence of YFP-ARHGAP36. Lysates were subjected to Flag IP. Normalized heavy/light ratio plot of the peptide evidences matching PRKAC. Only one peptide, highlighted in red, was upregulated in the presence of ARHGAP36 and was the only ubiquitylated PRKAC peptide identified. ( c ) The same as in a except where indicated PKAC-K285R-YFP was transfected and a Flag-Cherry control for Flag-ARHGAP36 was used. ( d ) The same as in c except lysates were subjected to His IP and immunoblotted with anti-GFP. ( e ) HEK293T cells were transfected with PKAC-Flag and CFP-ARHGAP36, the RRV mutant or CFP-Cherry control. Lysates were immunoblotted with the indicated antibodies. ( f ) Confocal micrographs of MDCK cells transfected with PKAC-YFP or PKAC-K285R-YFP and Flag-ARHGAP36. Twelve hours after transfection, cells were pretreated for 30 min with bafilomycin (100 nM), before cycloheximide (CHX; 1 mug ml -1 ) addition for a further 5 h. Cells were fixed and subjected to immunofluorescence using anti-GFP and anti-Flag. Scale bars, 10 mum. ( g ) HEK293T cells were transfected with PKAC-YFP in the presence or absence of Flag-ARHGAP36 and subjected to GFP IP followed by SRM-MS-b

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 8 ARHGAP36 is expressed in neuroblastoma cells and promotes aberrant activation of the Hedgehog pathway. ( a ) An amount of 10 mug lysate of the indicated cell lines was immunoblotted with the indicated antibodies. ARHGAP36 is only present in NGP and CLB-GA neuroblastoma cells. The predominant isoform seems to be isoform 3, expected size 46 kDa. ( b ) Histogram displays the distribution of the relative abundance of all measured proteins ( n =4448). ARHGAP36 and PKAC are within the same bin (pink). Bar plot shows the log 10 abundance of the indicated proteins relative to ARHGAP36, GAPDH (437-fold) and PKAC (0.95-fold). PKAC is the summed total of PRKACA and PRKACB. ( c ) NGP cell lysates were immunoprecipitated with an ARHGAP36 antibody or IgG rabbit control, and immunoblotted for ARHGAP36 or PKAC. 75% less lysate and eluate was run for ARHGAP36. ( d ) Confocal micrographs of NGP or CLB-GA cells transfected with GFP-Rab5-QL. Fixed cells were subjected to immunofluorescence using GFP, ARHGAP36 and PKAC antibodies. Scale bar, 5 mum. ( e ) Representative immunoblot of NGP cells treated with SMARTpool short interfering RNA (siRNA) against ARHGAP36 (si36) for 24 h. Cells were stimulated with 10 muM Forskolin and 100 muM IBMX for 30 min before collecting. Lysates were immunoblotted with the indicated antibodies. Lipo: reagent only control. NT1: non-targeting oligo control. Bands were densitometrically evaluated, normalized first to tubulin then to Lipo. Mean of five independe