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ANTIBODY INFORMATION VALIDATION SUMMARY IMMUNOCYTOCHEMISTRY IMMUNOHISTOCHEMISTRY WESTERN BLOT PROTEIN ARRAY ANTIGEN INFORMATION RELEVANT PUBLICATIONS

ANTIBODY INFORMATION

 
 

Antibody HPA024120

 

Antibody HPA050441

 

Antibody CAB004601

 

Provider

Atlas Antibodies
Sigma-Aldrich
  Atlas Antibodies
Sigma-Aldrich
  Santa Cruz Biotechnology
 

Product name

HPA024120   HPA050441   sc-25330  

Host species

Rabbit   Rabbit   Mouse  

Clonality

pAb   pAb   mAb  

Purity

Affinity purified using the PrEST-antigen as affinity ligand   Affinity purified using the PrEST-antigen as affinity ligand   Protein A/G  

Other gene match

     

Released in version

5   12   2  

References

1      

Proper citation

Atlas Antibodies Cat#HPA024120, RRID:AB_1858191   Atlas Antibodies Cat#HPA050441, RRID:AB_2681127   Santa Cruz Biotechnology Cat#sc-25330, RRID:AB_628384  

VALIDATION SUMMARY

 
 

Antibody HPA024120

 

Antibody HPA050441

 

Antibody CAB004601

 
Immunocytochemistry


Validated
  Supported   Supported  
Immunohistochemistry

Supported     Supported  
Western blot


Validated
  Supported   Supported  
Protein array

Supported   Supported    

IMMUNOCYTOCHEMISTRY

 
 

Antibody HPA024120

 

Antibody HPA050441

 

Antibody CAB004601

 

Formal validation: Genetic

Validated

Significant downregulation > 25% by both siRNA:s.
     

     

Figure description

Nuclear region of segmented cells in 10x-images      

Antibody dilution

1:105      
 

Standard validation

Supported   Supported   Supported  

     

Figure description

Immunofluorescent staining of human cell line U-251 MG shows localization to nucleoplasm.   Immunofluorescent staining of human cell line A-431 shows localization to nucleus & cytosol.   Immunofluorescent staining of human cell line A-431 shows localization to nucleus.  

Antibody dilution

1:100   1:64   1:25  

Literature conformity

The subcellular location is supported by literature.   The subcellular location is supported by literature.   The subcellular location is supported by literature.  

IMMUNOHISTOCHEMISTRY

 
 

Antibody HPA024120

 

Antibody HPA050441

 

Antibody CAB004601

 

Standard validation

Supported     Supported  

     

Figure description

Immunohistochemical staining of human pancreas shows moderate nuclear positivity.     Immunohistochemical staining of human pancreas shows strong nuclear positivity.  

Expression

RNA: detected in 37 tissues
Protein: detected in 77 cell types
    RNA: detected in 37 tissues
Protein: detected in 78 cell types
 

Retrieval

HIER pH6     HIER pH6  

Antibody dilution

1:300     1:100  

Literature conformity

Consistent with extensive gene/protein characterization data.     Consistent with extensive gene/protein characterization data.  

RNA consistency

Consistent with RNA expression data.     Consistent with RNA expression data.  

WESTERN BLOT

 
 

Antibody HPA024120

 

Antibody HPA050441

 

Antibody CAB004601

 

Formal validation: Genetic

Validated

Downregulation visible in both siRNA lanes
     

     

Figure description

Lane 1: Marker [kDa] 250, 130, 95, 72, 55, 36, 28, 17, 10
Lane 2: siRNA 1
Lane 3: siRNA 2
Lane 4: Scrambled
     

Target mass (kDa)

183.3, 182.7      

Loading control

Total protein image      

Antibody dilution

1:233      
 

Standard validation

Supported

Analysis performed using a standard panel of samples.
Band of predicted size in kDa (+/-20%) with additional bands present.
  Supported

Analysis performed using a standard panel of samples.
Band of predicted size in kDa (+/-20%) with additional bands present.
  Supported

Analysis performed using a standard panel of samples.
Single band corresponding to the predicted size in kDa (+/-20%).
 

     

Figure description

Lane 1: Marker [kDa] 230, 130, 95, 72, 56, 36, 28, 17, 11
Lane 2: RT4
Lane 3: U-251 MG
Lane 4: Human Plasma
Lane 5: Liver
Lane 6: Tonsil
  Lane 1: Marker [kDa] 250, 130, 95, 72, 55, 36, 28, 17, 10
Lane 2: RT4
Lane 3: U-251 MG
Lane 4: Human Plasma
Lane 5: Liver
Lane 6: Tonsil
  Lane 1: Marker [kDa] 230, 110, 82, 49.3, 32.2, 25.5, 17.6
Lane 2: RT4
Lane 3: U-251 MG
Lane 4: Human Plasma
Lane 5: Liver
Lane 6: Tonsil
 

Target mass (kDa)

183.3, 182.7   183.3, 182.7   183.3, 182.7, 130.5  

Antibody dilution

1:250   1:320   1:500  

PROTEIN ARRAY

 
 

Antibody HPA024120

 

Antibody HPA050441

 

Antibody CAB004601

 

Standard validation

Supported

Pass with single peak corresponding to interaction only with its own antigen.
  Supported

Pass with single peak corresponding to interaction only with its own antigen.
   

     

Figure description

Antibody specificity analysis with protein arrays. Predicted and matching interactions are shown in green.   Antibody specificity analysis with protein arrays. Predicted and matching interactions are shown in green.    

Antibody dilution

1:3000   1:4000    

ANTIGEN INFORMATION

 
 

Antibody HPA024120

 

Antibody HPA050441

 

Antibody CAB004601

 

Antigen

Recombinant protein fragment   Recombinant protein fragment   Recombinant protein  

Length (aa)

143   138    

Antigen sequence

LWKEDLAAFVEELDKVESQEREDVLAGMSGKAIKGKVGKPKVKKLQLEET
MPSPYGRRIIPEITAMKADASKKLLKKKKGDLDTAAVKVEFDEEFSGAPV
EGAGEEALTPSVPINKGPKPKREKKEPGTRVRKTPTSSGKPSA
 
DKVPSKTVAAKKGKPSSDTVPKPKRAPKQKKVVEAVNSDSDSEFGIPKKT
TTPKGKGRGAKKRKASGSENEGDYNPGRKTSKTTSKKPKKTSFDQDSDVD
IFPSDFPTEPPSLPRTGRARKEVKYFAESDEEEDDVDF
   

Matching transcripts

TOP2B-001 - ENSP00000396704 [100%]
TOP2B-201 - ENSP00000264331 [100%]
  TOP2B-001 - ENSP00000396704 [100%]
TOP2B-201 - ENSP00000264331 [100%]
   

Other gene match

     
ANTIGEN VIEW
TOP2B-001
TOP2B-201

RELEVANT PUBLICATIONS

 
 

Antibody HPA024120

 

Antibody HPA050441

 

Antibody CAB004601

 

1.

Uncovering Hidden Layers of Cell Cycle Regulation through Integrative Multi-omic Analysis
Aviner R et al
PLoS Genet 2015;11(10):e1005554
     
 

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