PA5-21117

antibody from Invitrogen Antibodies

Targeting: RABGEF1 rabex-5, RABEX5

Western blot (Data presented on provider website)

Immunohistochemistry (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-21117

Provider

Invitrogen Antibodies

Product name

Rabex-5 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

A suggested positive control is rat brain tissue lysate. PA5-21117 can be used with blocking peptide PEP-1231.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

1 mg/mL

Storage

Maintain refrigerated at 2-8°C for up to 3 months. For long term storage store at -20°C

References

Submitted references

Critical role for Piccolo in synaptic vesicle retrieval.

Ackermann F, Schink KO, Bruns C, Izsvák Z, Hamra FK, Rosenmund C, Garner CC
eLife 2019 May 10;8

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 5--figure supplement 1. Synaptic Rabex5 levels as well as Rabex5 amounts at PI3P-positive organelles are reduced in neurons lacking Piccolo. ( A ) Immunocytochemical staining of hippocampal neurons for Rabex5. ( B ) Quantification of ( A ). Rabex5 is present at Pclo wt/wt and Pclo gt/gt synapses, no difference in the intensities is detectable ( Pclo wt/wt = 1 +- 0.02, n = 2397 synapses; Pclo gt/gt = 0.96 +- 0.02, n = 1802 synapses; seven independent experiments). ( C ) Images depicting Rabex5 intensities at PI3P-positive organelles. ( D ) Quantification of ( C ). Less Rabex5 is present at PI3P-positive membranes in Pclo gt/gt neurons ( Pclo wt/wt = 1 +- 0.04, n = 728 puncta; Pclo gt/gt = 0.80 +- 0.03, n = 652 puncta; six independent experiments). ( E ) Quantification of double positive compartments along axons. The fraction of GFP-2x-FYVE/Rabex5 double positive vesicles is increased in Pclo gt/gt neurons ( Pclo gt/gt = 1.49 +- 0.33, n = 5 independent experiments). ( F ) The relative percentage of GFP-2x-FYVE-Rabex5-Rab5 triple positive compartments is decreased in Pclo gt/gt neurons ( Pclo gt/gt = 0.57 +- 0.14, n = 5 independent experiments). Scale bar represents 10 mum. Error bars in bar graph represent 95% confidence intervals. Numbers given represent mean +- SEM, Student`s t-test.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 8--figure supplement 1. Silencing synaptic activity affects synaptic levels of endosome proteins in Pclo wt/wt and Pclo gt/gt neurons. ( A and B ) Images of Pclo wt/wt ( A ) and Pclo gt/gt ( B ) neurons stained for Synaptophysin and Rab5 with and without TTX treatment. ( C ) Quantification of ( A and B ). Synaptic Rab5 levels do not significantly alter in Pclo wt/wt and Pclo gt/gt synapses upon TTX treatment ( Pclo wt/wt = 1 +- 0.05, n = 391 synapses; Pclo wt/wt (TTX) = 0.73 +- 0.05, n = 482 synapses; Pclo gt/gt = 1.14 +- 0.09, n = 400 synapses; Pclo gt/gt (TTX) = 0.90 +- 0.04, n = 372 synapses; two independent experiments). ( D and E ) Images of Pclo wt/wt ( D ) and Pclo gt/gt ( E ) neurons stained for Synaptophysin and EEA1 with and without TTX treatment. ( F ) Quantification of ( D and E ). In Pclo wt/wt synapses EEA1 levels drop upon TTX treatment ( Pclo wt/wt = 1 +- 0.03, n = 1653 synapse; Pclo wt/wt (TTX) = 0.72 +- 0.03, n = 1887 synapses; four independent experiments). In contrast, EEA1 levels slightly increase in Pclo gt/gt synapses due to TTX treatment ( Pclo gt/gt = 0.68 +- 0.04, n = 918; Pclo gt/gt (TTX) = 0.78 +- 0.03, n = 1352; four independent experiments). ( G and H ) Pclo wt/wt ( G ) and Pclo gt/gt ( H ) neurons stained for Synaptophysin and Rabex5 with and without TTX treatment. ( I ) Quantification of ( G and H ). Rabex5 level at synapses slightly decrease in Pclo wt/wt and Pclo gt/gt neurons upon TTX trea