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MCM4
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ANTIBODY INFORMATION VALIDATION SUMMARY IMMUNOCYTOCHEMISTRY IMMUNOHISTOCHEMISTRY WESTERN BLOT PROTEIN ARRAY ANTIGEN INFORMATION RELEVANT PUBLICATIONS

ANTIBODY INFORMATION

 
 

Antibody HPA004873

 

Antibody HPA031052

 

Antibody CAB004497

 

Provider

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

Product name

HPA004873   HPA031052   sc-28317  

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

3   12   3  

References

     

Proper citation

Atlas Antibodies Cat#HPA004873, RRID:AB_1079337   Atlas Antibodies Cat#HPA031052, RRID:AB_2673712   Santa Cruz Biotechnology Cat#sc-28317, RRID:AB_627916  

VALIDATION SUMMARY

 
 

Antibody HPA004873

 

Antibody HPA031052

 

Antibody CAB004497

 
Immunocytochemistry


Validated

Validated
 

Validated

Validated
   
Immunohistochemistry


Validated
    Supported  
Western blot


Validated
 

Validated
  Supported  
Protein array

Supported   Supported    

IMMUNOCYTOCHEMISTRY

 
 

Antibody HPA004873

 

Antibody HPA031052

 

Antibody CAB004497

 

Formal validation: Genetic

Validated

Significant downregulation > 25% by one siRNA.
 
Validated

Significant downregulation > 25% by both siRNA:s.
   

 
   

Figure description

Nuclear region of segmented cells in 10x-images   Nuclear region of segmented cells in 10x-images    

Antibody dilution

1:10   1:101    
 

Formal validation: Independent

Validated

Antibody staining overlaps with antibody HPA031052.
 
Validated

Antibody staining overlaps with antibody HPA004873.
   
 

Standard validation

Supported   Supported    

     

Figure description

Immunofluorescent staining of human cell line U-2 OS shows localization to nucleoplasm.   Immunofluorescent staining of human cell line U-2 OS shows localization to nucleoplasm.    

Antibody dilution

1:18   1:101    

Literature conformity

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

IMMUNOHISTOCHEMISTRY

 
 

Antibody HPA004873

 

Antibody HPA031052

 

Antibody CAB004497

 

Formal validation: Orthogonal

Validated

Spearman correlation >0.6 for protein and RNA expression in tissues.
     

     

Figure description

Distribution of protein expression (antibody staining) and RNA expression (TPM) across 36 tissues.      
 

Standard validation

Supported     Supported  

     

Figure description

Immunohistochemical staining of human lymph node shows strong nuclear positivity in cells in reaction center.     Immunohistochemical staining of human lymph node shows strong nuclear positivity in reaction center cells  

Expression

RNA: detected in 37 tissues
Protein: detected in 48 cell types
    RNA: detected in 37 tissues
Protein: detected in 73 cell types
 

Retrieval

HIER pH6     HIER pH6  

Antibody dilution

1:50     1:100  

Literature conformity

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

RNA consistency

Mainly consistent with RNA expression data.     Consistent with RNA expression data.  

WESTERN BLOT

 
 

Antibody HPA004873

 

Antibody HPA031052

 

Antibody CAB004497

 

Formal validation: Genetic

Validated

Downregulation visible in both siRNA lanes
 
Validated

Downregulation visible in both siRNA lanes
   

     

Figure description

Lane 1: Marker [kDa] 250, 150, 100, 75, 50, 37, 25, 20, 15, 10
Lane 2: siRNA 1
Lane 3: siRNA 2
Lane 4: Scrambled
  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)

96.6   96.6, 23.5    

Loading control

Total protein image   Total protein image    

Antibody dilution

1:100   1:224    
 

Standard validation

Supported

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

Analysis performed using a standard panel of samples.
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.
 

     

Figure description

Lane 1: Marker [kDa] 220, 112, 84, 47, 32, 26, 16.8
Lane 2: RT4
Lane 3: U-251 MG
Lane 4: Human Plasma
Lane 5: Liver
Lane 6: Tonsil
  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] 229, 112, 83.5, 47.9, 32.3, 26.5, 17.2
Lane 2: RT4
Lane 3: U-251 MG
Lane 4: Human Plasma
Lane 5: Liver
Lane 6: Tonsil
 

Target mass (kDa)

96.6   96.6, 23.5   96.6, 31.7, 23.5, 14.8  

Antibody dilution

1:250   1:250   1:500  

PROTEIN ARRAY

 
 

Antibody HPA004873

 

Antibody HPA031052

 

Antibody CAB004497

 

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

1:2000   1:3000    

ANTIGEN INFORMATION

 
 

Antibody HPA004873

 

Antibody HPA031052

 

Antibody CAB004497

 

Antigen

Recombinant protein fragment   Recombinant protein fragment   Recombinant protein  

Length (aa)

139   116    

Antigen sequence

LPHTLLSRFDLIFLMLDPQDEAYDRRLAHHLVALYYQSEEQAEEELLDMA
VLKDYIAYAHSTIMPRLSEEASQALIEAYVDMRKIGSSRGMVSAYPRQLE
SLIRLAEAHAKVRLSNKVEAIDVEEAKRLHREALKQSAT
 
SSPPQMHSSAIPLDFDVSSPLTYGTPSSRVEGTPRSGVRGTPVRQRPDLG
SAQKGLQVDLQSDGAAAEDIVASEQSLGQKLVIWGTDVNVAACKENFQRF
LQRFIDPLAKEEENVG
   

Matching transcripts

MCM4-001 - ENSP00000262105 [99%]
MCM4-004 - ENSP00000430194 [99%]
  MCM4-001 - ENSP00000262105 [100%]
MCM4-004 - ENSP00000430194 [100%]
MCM4-009 - ENSP00000428833 [100%]
   

Other gene match

     
ANTIGEN VIEW
MCM4-001
MCM4-004
MCM4-009
 

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