Rab 5

GTP-binding protein involved in endocytosis

Fact Sheet

Monoclonal mouse antibody, clone 621.3
Cat. No. 108 011 IgG, unlabeled	100 µg purified IgG, lyophilized. For reconstitution add 100 µl H₂O to get a 1mg/ml solution of antibody in PBS. Then aliquot and store at -20°C until use.
Applications	WB: 1 : 1000 up to 1 : 10000 (AP staining) blot IP: yes ICC: 1 : 100 up to 1 : 1000 (see remarks) IHC: not tested yet
Clone	621.3
Subtype	IgG2a
Immunogen	Recombinant full length rat rab 5a.
Reactivity	Human, mouse, rat, hamster; other species not tested yet.
Specificity	Specific for rab 5 (probably only rab 5a). No cross-reactivity to other rab proteins.
Remarks	ICC: Not suitable for ICC of aldehyde- fixed tissues or cells. Recommended for human samples. This antibody was used very successfully for immunoisolation of early endosomes and for the differentiation of early endosomes from related trafficking organelles in neurons and nonneuronal cells.

Monoclonal mouse antibody, clone 621.4
Cat. No. 108 101 ascites	100 µl ascites, lyophilized. For reconstitution add 100 µl H₂O, then aliquot and store at -20°C until use.
Applications	WB: 1 : 1000 up to 1 : 10000 (AP staining) IP: yes ICC: 1 : 100 up to 1 : 1000 (but see remarks) IHC: not tested yet
Clone	621.4
Subtype	IgG2a
Immunogen	Recombinant full length rat rab 5a.
Reactivity	Human, rat, mouse; other species not tested.
Specificity	Specific for rab 5 (probably only rab 5a). No cross-reactivity to other rab proteins.
Remarks	ICC: Not suitable for ICC of aldehyde- fixed tissues or cells. This antibody was used very successfully for immunoisolation of early endosomes and for the differentiation of early endosomes from related trafficking organelles in neurons and nonneuronal cells.

Polyclonal rabbit antibody
Cat. No. 108 002 crude antiserum	200 µl antiserum, lyophilized. For reconstitution add 200 µl H₂O, then aliquot and store at -20°C until use.
Applications	WB: 1 : 1000 (AP staining) blot IP: not tested yet ICC: not tested yet IHC: not tested yet
Immunogen	Synthetic peptide (aa 182-196 in mouse rab 5) coupled to key-hole limpet hemocyanin via an added N-terminal cysteine residue.
Reactivity	Human, rat, mouse; other species not tested yet.
Specificity	Specific for Rab 5.

Control peptide for rabbit antibody
Cat. No. 108-0P	100 µg lyophilized peptide. For reconstitution add 100 µl H₂O to get a 1mg/ml solution in PBS. Then aliquot and store at -20°C until use. Control peptides should also be stored at -20°C when still lyophilized!
Recommended dilution	Optimal concentrations should be determined by the end-user.
Remarks	This control peptide consists of the synthetic peptide (aa 182-196 in mouse rab 5) that has been used for immunization. It has been tested in preadsorption experiments and blocks efficiently and specifically the corresponding signal in Western blots. The amount of peptide needed for efficient blocking depends on the titer and on the affinity of the antibody to the antigen.