RELB antibody (AA 1-579)

Catalog No. ABIN519730

Product Details anti-RELB Antibody

Target: RELB (V-Rel Reticuloendotheliosis Viral Oncogene Homolog B (RELB))

Binding Specificity: AA 1-579

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This RELB antibody is un-conjugated

Application: Western Blotting (WB), Proximity Ligation Assay (PLA)

Purpose: Rabbit polyclonal antibody raised against a full-length human RELB protein.

Sequence: MLRSGPASGP SVPTGRAMPS RRVARPPAAP ELGALGSPDL SSLSLAVSRS TDELEIIDEY IKENGFGLDG GQPGPGEGLP RLVSRGAASL STVTLGPVAP PATPPPWGCP LGRLVSPAPG PGPQPHLVIT EQPKQRGMRF RYECEGRSAG SILGESSTEA SKTLPAIELR DCGGLREVEV TACLVWKDWP HRVHPHSLVG KDCTDGICRV RLRPHVSPRH SFNNLGIQCV RKKEIEAAIE RKIQLGIDPY NAGSLKNHQE VDMNVVRICF QASYRDQQGQ MRRMDPVLSE PVYDKKSTNT SELRICRINK ESGPCTGGEE LYLLCDKVQK EDISVVFSRA SWEGRADFSQ ADVHRQIAIV FKTPPYEDLE IVEPVTVNVF LQRLTDGVCS EPLPFTYLPR DHDSYGVDKK RKRGMPDVLG ELNSSDPHGI ESKRRKKKPA ILDHFLPNHG SGPFLPPSAL LPDPDFFSGT VSLPGLEPPG GPDLLDDGFA YDPTAPTLFT MLDLLPPAPP HASAVVCSGG AGAVVGETPG PEPLTLDSYQ APGPGDGGTA SLVGSNMFPN HYREAAFGGG LLSPGPEAT

Cross-Reactivity: Human

Characteristics: Antibody reactive against mammalian transfected lysate.

Immunogen: RELB (NP_006500.2, 1 a.a. ~ 579 a.a) full-length human protein.

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Restrictions: For Research Use only

Independent Validation (IF)

Validation #104076 (Immunofluorescence)

by: Klinik für Anästhesiologie, Intensivmedizin und Schmerztherapie des Universitätsklinikums Knappschaftskrankenhaus Bochum

No.: #104076

Date: 11/21/2019

Antigen: RELB

Lot Number: 09098

Method validated: Immunofluorescence

Positive Control:

U-937 human monocytic cell line

Negative Control:

No primary antibody control

Notes:

Passed. ABIN519730 specifically stains RelB in U-937 human monocytic cells.

Validation Images

U937 monocytic cells stained with with anti-RelB antibody ABIN519730 and a TRITC-labeled secondary antibody (bottom row, red). Counterstain with DAPI (blue). Negative control without primary antibody (top row) shows no specific TRITC signal.

Full Methods

Primary Antibody: ABIN519730

Secondary Antibody: TRITC-conjugated AffiniPure donkey anti-rabbit IgG antibody (611-025-215, Jackson ImmunoResearch Europe Ltd.)

Full Protocol:

• Grow U-937 cells in RPMI 1640 Medium (Gibco, 21875034) supplemented with 10 % FCS (Gibco, A3840001) and Pen/Strep (Gibco, 15140122), at 37°C and 5% CO₂ in 300µL in 24-well plates (Sarstedt AG&Co. KG).
• Use cytospin to spin cells onto slide.
• Fix cells on coverslide in 4% paraformaldehyde (Carl Roth GmbH, 0335.1) for 30min at RT.
• Wash cells 3x for 5min with PBS (Sigma-Aldrich, 8537).
• Permeabilize cells in 100µL PBS containing 0.1 % Triton X-100 (Sigma-Aldrich, T8787) for 5min at RT.
• Wash cells 3x for 5min with PBS.
• Block non-specific binding with 1x Duolink Blocking solution (Sigma-Aldrich, DUO82007) for 30min at RT.
• Incubate cells with primary rabbit anti-human RelB antibody (antibodies-online, ABIN519730, lot 09098 WUlz) diluted 1:100 in 1x Duolink Antibody diluent (Sigma-Aldrich, DUO82008) ON at 4°C.
• Wash cells 3x for 5min with PBS.
• Incubate cells with secondary TRITC-conjugated AffiniPure donkey anti-rabbit IgG antibody (611-025-215, Jackson ImmunoResearch Europe Ltd.) diluted 1:400 in 1x Duolink Antibody diluent for 1 h at RT in the dark.
• Wash cells 2x for 5min with PBS.
• Mount coverslips on glass slides in 50µL ProLong Gold antifade mountant with DAPI (Invitrogen, P36931).
• Image acquisition with an Olympus widefield microscope.

Experimental Notes:

Strong red staining depending on the presence of primary antibody indicates that the antibody works. Negative control lacking the primary antibody shows only background fluorescence.

Handling

Buffer: In 1x PBS, pH 7.4

Handling Advice: Aliquot to avoid repeated freezing and thawing.

Storage: -20 °C

Storage Comment: Store at -20°C or lower. Aliquot to avoid repeated freezing and thawing.

Target Details for RELB

Target: RELB (V-Rel Reticuloendotheliosis Viral Oncogene Homolog B (RELB))

Alternative Name: RELB (RELB Products)

Synonyms: shep antibody, I-REL antibody, IREL antibody, REL-B antibody, XRelB antibody, relb-A antibody, GC-rich antibody, avian reticuloendotheliosis viral (v-rel) oncogene related B antibody, RELB proto-oncogene, NF-kB subunit antibody, v-rel avian reticuloendotheliosis viral oncogene homolog B S homeolog antibody, Relb antibody, RELB antibody, relb.S antibody

Background: Full Gene Name: v-rel reticuloendotheliosis viral oncogene homolog B
Synonyms: I-REL,IREL

Gene ID: 5971

NCBI Accession: NM_006509

Pathways: NF-kappaB Signaling, RTK Signaling