PA5-28522

antibody from Invitrogen Antibodies

Targeting: SPOP BTBD32, TEF2

Provider product page for PA5-28522

Western blot

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [2]
Comments [0]
Validations
Western blot [4]
Other assay [3]

Submit

Product number: PA5-28522 - Provider product page
Provider: Invitrogen Antibodies
Product name: SPOP Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant protein fragment
Description: Recommended positive controls: HepG2, HepG2 nuclear extract, Mouse brain, SPOP-transfected 293T. Predicted reactivity: Mouse (100%), Rat (100%), Zebrafish (99%), Xenopus laevis (100%), Rhesus Monkey (100%), Bovine (100%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
Reactivity: Human, Mouse
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 0.1 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

SPOP protein structure - PA5-28522 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of SPOP using 30 µg of 293T lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with a SPOP polyclonal antibody (Product # PA5-28522) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of SPOP was performed by separating 30 µg of HepG2 whole cell and nuclear extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a SPOP Polyclonal Antibody (Product # PA5-28522) at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using SPOP Polyclonal Antibody (Product # PA5-28522). Sample (50 µg of whole cell lysate). Lane A: mouse brain. 10% SDS PAGE. SPOP Polyclonal Antibody (Product # PA5-28522) diluted at 1:2,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of SPOP was performed by separating 30 µg of non-transfected (–) and transfected (+) 293T whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a SPOP Polyclonal Antibody (Product # PA5-28522) at a dilution of 1:5000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4. Loss of Tbx3 results in Gli3 protein instability and aberrant localization of Kif7 in limb bud cilia. ( A ) Representative immunoblot (N=3) blot of E10.75 forelimb bud lysates prepared from microdissected Tbx3 fl/+ control anterior limb (CA), Tbx3;PrxCre mutant anterior limb (MA), control posterior (CP), and mutant posterior (MP) probed for Gli3 and betatubulin loading control. Note decreased level of Gli3FL and Gli3R, and multiple bands of lower molecular weight than Gli3R in MA sample. Densitometry of Gli3R bands in red box in N revealed that in this representative experiment, the level of Gli3R was 7.4 fold decreased in mutant anterior relative to control anterior. ( A' ) Longer exposure of top of blot shown in panel A to examine Gli3FL band. The control (CA) Gli3FL band is 31 fold more intense than mutant (MA, virtually undetectable). ( B ) Immunoblot of lysates from E10.5 forelimb buds immunoprecipitated (IB) with antibodies listed at top and immunoblotted (IB) for Tbx3. Lane 5 shows that immunoprecipitation with anti-Kif7 antibody co-IPs Tbx3. ( C ) As in panel B, but assayed for Kif7. ( D ) Co-IP assay of Myc-tagged Tbx3 and Flag-tagged GFP overexpressed in HEK293 cells. IP was performed with antibodies listed at top and immunoblotted for Tbx3. Myc-tagged Tbx3 co-IPs with Flag-tagged Kif7. Input lane was 5 s exposure (5"" exp) while other lanes were 15 s (15"" exp). ( E ) As in D , but in this case, blot probed for Kif7; confirms interaction of tagged, o

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7--figure supplement 3. Tbx3 does not co-IP with Sufu or Spop in mouse embryo lysates. ( A , B ) Immunoprecipitations/Immunoblot assaying for interaction between endogenous Tbx3 and Sufu in E10.5 mouse embryo lysates. Sufu did not co-IP Tbx3 ( A ) nor did Tbx3 co-IP Sufu ( B ). ( C ) Immunoprecipitations/Immunoblot assaying for interaction between Tbx3 and Spop in control and Tbx3 Deltafl/Deltafl E10.5 mouse embryo lysates. No interaction was detected. Note increased Spop in mutant, consistent with previous result in Figure 4J and data in Figure 8--figure supplement 1 . DOI: http://dx.doi.org/

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 8--figure supplement 1. Altered protein levels observed in mutant limb buds are also apparent in whole embryos. ( A , B ) Immunoblots on protein lysates prepared from E10.5 forelimb buds ( A ) and whole embryos ( B ). Actin is loading control. ( C ) Quantitation of protein levels in A and B comparing amount detected in control to mutant. Note increased levels of Sufu and Spop in mutant limbs and embryos that results in ratios of control to mutant