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Method

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Antibodies

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

ANTIBODY INFORMATION

 
 

Antibody HPA019799

 

Antibody HPA061282

 

Antibody CAB016281

 

Provider

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

Product name

HPA019799   HPA061282   sc-32311  

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   13   4  

References

1      

VALIDATION SUMMARY

 
 

Antibody HPA019799

 

Antibody HPA061282

 

Antibody CAB016281

 
Immunocytochemistry


Validated
  Approved   Approved  
Immunohistochemistry

Supported     Supported  
Western blot


Validated
  Supported   Supported  
Protein array

Supported   Supported    

IMMUNOCYTOCHEMISTRY

 
 

Antibody HPA019799

 

Antibody HPA061282

 

Antibody CAB016281

 

Formal validation: Genetic

Validated

Significant downregulation > 25% by one siRNA.
     

 

Relative fluorescence intensity (RFI)

     

Figure description

Nuclear region of segmented cells in 10x-images      

Antibody dilution

1:100      
 

Standard validation

Supported   Approved   Approved  

     

Figure description

Immunofluorescent staining of human cell line U-2 OS shows localization to nuclear speckles.   Immunofluorescent staining of human cell line U-2 OS shows localization to nuclear speckles.   Immunofluorescent staining of human cell line U-2 OS shows localization to nucleus & nucleoli.  

Antibody dilution

1:100   1:27   1:2000  

Literature conformity

The subcellular location is supported by literature.   The subcellular location is partly supported by literature or no literature is available.   The subcellular location is partly supported by literature or no literature is available.  

IMMUNOHISTOCHEMISTRY

 
 

Antibody HPA019799

 

Antibody HPA061282

 

Antibody CAB016281

 

Standard validation

Supported     Supported  

     

Figure description

Immunohistochemical staining of human testis shows strong nuclear positivity in cells in seminiferus tubules.     Immunohistochemical staining of human gallbladder shows nuclear positivity in glandular cells.  

Expression

RNA: detected in 37 tissues
Protein: detected in 80 cell types
    RNA: detected in 37 tissues
Protein: detected in 78 cell types
 

Retrieval

HIER pH6     HIER pH6  

Antibody dilution

1:225     1:5000  

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 HPA019799

 

Antibody HPA061282

 

Antibody CAB016281

 

Formal validation: Genetic

Validated

Downregulation visible in one of two 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)

27.6      

Loading control

Total protein image      

Antibody dilution

1:222      
 

Standard validation

Supported

Single band corresponding to the predicted size in kDa (+/-20%).
  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] 250, 130, 95, 72, 55, 36, 28, 17, 10
Lane 2: Negative control (vector only transfected HEK293T lysate)
Lane 3: Over-expression Lysate (Co-expressed with a C-terminal myc-DDK tag (~3.1 kDa) in mammalian HEK293T cells, LY417070)
  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] 250, 130, 95, 72, 55, 36, 28, 17, 11
Lane 2: RT4
Lane 3: U-251 MG
Lane 4: Human Plasma
Lane 5: Liver
Lane 6: Tonsil
 

Target mass (kDa)

27.6   27.6   27.6  

Antibody dilution

1:250   1:90   1:500  

PROTEIN ARRAY

 
 

Antibody HPA019799

 

Antibody HPA061282

 

Antibody CAB016281

 

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   1:1000    

ANTIGEN INFORMATION

 
 

Antibody HPA019799

 

Antibody HPA061282

 

Antibody CAB016281

 

Antigen

Recombinant protein fragment   Recombinant protein fragment   Recombinant protein  

Length (aa)

128   37    

Antigen sequence

EFGTLKKAAVHYDRSGRSLGTADVHFERKADALKAMKQYNGVPLDGRPMN
IQLVTSQIDAQRRPAQSVNRGGMTRNRGAGGFGGGGGTRRGTRGGARGRG
RGAGRNSKQQLSAEELDAQLDAYNARMD
 
GGGRNRPAPYSRPKQLPDKWQHDLFDSGFGGGAGVET
   

Matching transcripts

ALYREF-001 - ENSP00000421592 [100%]
  ALYREF-001 - ENSP00000421592 [100%]
   

Other gene match

     
ANTIGEN VIEW
ALYREF-001

RELEVANT PUBLICATIONS

 
 

Antibody HPA019799

 

Antibody HPA061282

 

Antibody CAB016281

 

1.

Differential expression of THOC1 and ALY mRNP biogenesis/export factors in human cancers
Domínguez-Sánchez MS et al
BMC Cancer 2011;11:77
     
 

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