PA5-95195

antibody from Invitrogen Antibodies

Targeting: UBB FLJ25987, MGC8385

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-95195

Provider

Invitrogen Antibodies

Product name

Ubiquitin B Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL. Positive Control - WB: human Caco-2 whole cell, human K562 whole cell, human THP-1 whole cell, rat PC-12 whole cell, rat brain tissue, mouse brain tissue. IHC: rat intestine tissue, human mammary cancer tissue, mouse intestine tissue. ICC/IF: A431 cell.|Store at -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freeze-thaw cycles.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

500 µg/mL

Storage

-20°C

References

Submitted references

Endogenous Fluorescent Proteins in the Mucus of an Intertidal Polychaeta: Clues for Biotechnology.

Rodrigo AP, Lopes A, Pereira R, Anjo SI, Manadas B, Grosso AR, Baptista PV, Fernandes AR, Costa PM
Marine drugs 2022 Mar 25;20(4)

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Ubiquitin in, Lane 1: human Caco-2 whole cell lysates, Lane 2: human K562 whole cell lysates, Lane 3: human THP-1 whole cell lysates, Lane 4: rat PC-12 whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: mouse brain tissue lysates, . Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. The membrane was blocked with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with Ubiquitin B Polyclonal Antibody (Product # PA5-95195) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0. 1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for Ubiquitin at approximately 9KD. The expected band size for Ubiquitin is at 9KD.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of Ubiquitin using anti-Ubiquitin antibody (Product # PA5-95195) . Ubiquitin was detected in a section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 5μg/mL rabbit anti-Ubiquitin antibody (Product # PA5-95195) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of Ubiquitin using anti-Ubiquitin antibody (Product # PA5-95195) . Ubiquitin was detected in a section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 5μg/mL rabbit anti-Ubiquitin antibody (Product # PA5-95195) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Ubiquitin B in paraffin-embedded rat intestine tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with Ubiquitin B polyclonal antibody (Product # PA5-95195) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Ubiquitin B in paraffin-embedded human mammary cancer tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with Ubiquitin B polyclonal antibody (Product # PA5-95195) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Ubiquitin B in paraffin-embedded mouse intestine tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with Ubiquitin B polyclonal antibody (Product # PA5-95195) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Ubiquitin B in paraffin-embedded mouse intestine tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with Ubiquitin B polyclonal antibody (Product # PA5-95195) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical localization of ubiquitin B in Eulalia . ( A ) Alignment of peptides from MS/MS on fluorescent SDS-PAGE gel bands, RNAseq sequences with positive matches to ubiquitin, and the human ubiquitin B sequence against which the polyclonal antibody was used. The antibody was chosen based on similarity with Eulalia ubiquitin isoform 3. ( B ) Paraffin section across a parapodium of the worm fixed with Zenker and stained with HE (hematoxylin and eosin). Hemocytes are identified by the arrows. Inset: Paraffin section fixed with glutaraldehyde and stained with TC (tetrachrome stain) showing hepatocytes naturally colored by green pigments. ( C ) Compositive of a cryopreserved section of a parapodia marked for ubiquitin B. ( D ) Negative control of cryopreserved section of a parapodia. ( E ) Paraffin section fixed with Zenker and stained with HE; arrows are indicative of stem cells. ( F ) Composite of a paraffin section of the ventral lateral section marked for ubiquitin B. Arrows indicate the stem cells marked by the antibody. ( G ) Negative control of cryopreserved section of the ventral lateral area, close to the parapodia. ( H ) Paraffin section fixed with glutaraldehyde and stained with HE, showing the female gametes in the celomic cavity. ( I ) Composite of celomic cavity with positive signal for ubiquitin B (arrow). ( J ) Negative control of cryopreserved section of the celomic cavity.