PA1-41220

antibody from Invitrogen Antibodies

Targeting: RTN4 ASY, KIAA0886, NOGO, NSP-CL

Western blot (Data presented on provider website)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-41220

Provider

Invitrogen Antibodies

Product name

Nogo-A/B Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Suggested positive control: human, mouse or rat brain.

Reactivity

Human, Mouse, Rat, Rabbit

Host

Rabbit

Isotype

IgG

Vial size

200 µL

Concentration

1.0 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

REEP5 depletion causes sarco-endoplasmic reticulum vacuolization and cardiac functional defects.

Lee SH, Hadipour-Lakmehsari S, Murthy HR, Gibb N, Miyake T, Teng ACT, Cosme J, Yu JC, Moon M, Lim S, Wong V, Liu P, Billia F, Fernandez-Gonzalez R, Stagljar I, Sharma P, Kislinger T, Scott IC, Gramolini AO
Nature communications 2020 Feb 19;11(1):965

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Reticulon-4 was performed using 70% confluent log phase U-87 MG cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Nogo-A/B Polyclonal Antibody (Product # PA1-41220) at (1:100 dilution) in 0.1% BSA, incubated at 4-degree Celsius overnight, and labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 6 Mass spectrometry analysis identifies REEP5 interactions with known cardiac SR shaping proteins. a Immunoblot analysis of nickel-His purification of CnVA-REEP5 from transfected HEK293 cells shows stable expression of CnVA-REEP5 and successful immunoprecipitation of CnVA-REEP5 monomer (~34 kDa) and dimer (~68 kDa). b Identification of known SR/ER-shaping proteins as REEP5 interacting proteins by mass spectrometry analysis. Average precursor MS1 peak areas (peptide m/z signal) as defined by iBAQ (intensity based absolute quantification) are shown, n = 3 independent mass spectrometry runs. To calculate fold change, average values (n.d.--not detected) were inputted with a value of 10. RTN reticulon, ATL atlastin, CKAP4 cytoskeleton-associated protein 4, ACTC1 alpha cardiac muscle actin 1, MYL6 myosin light chain 6, GAPDH glyceraldehyde-3-phosphate dehydrogenase. Asterisks indicate a statistically significant p value in a Tukey's multiple comparison analysis where * p < 0.05. c RNASeq analysis of the RTN, ATL families of proteins, and CKAP4 in human fetal heart and adult heart tissue data using data from Human Protein Atlas. d Immunoblot analysis of nickel-His REEP5 immunoprecipitation lysates for RTN4/Nogo-A/B, ATL3, and CKAP4, n = 3. e HEK293 c e lls were transfected with myc-tagged Nogo-A and Nogo-B plasmids and detected with myc and alpha-tubulin antibodies. f Co-immunoprecipitation assay of cotransfected HEK293 cells with anti-REEP5 antibody (left panel) and anti-