RELB antibody (AA 1-579)

Catalog No. ABIN948396

Product Details anti-RELB Antibody

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

Binding Specificity: AA 1-579

Reactivity: Human

Host: Mouse

Clonality: Polyclonal

Conjugate: This RELB antibody is un-conjugated

Application: Western Blotting (WB), Immunofluorescence (IF)

Purpose: Mouse 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 #104077 (Immunofluorescence)

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

No.: #104077

Date: 11/21/2019

Antigen: RELB

Lot Number: 10307

Method validated: Immunofluorescence

Positive Control:

U-937 human monocytic cell line

Negative Control:

No primary antibody control

Notes:

Passed. ABIN948396 specifically stains RelB in U-937 human monocytic cells.

Validation Images

U937 monocytic cells stained with with anti-RelB antibody ABIN948396 and an AF488-labeled secondary antibody (bottom row, green). Counterstain with DAPI (blue). Negative control without primary antibody (top row) shows no AF488 signal.

Full Methods

Primary Antibody: ABIN948396

Secondary Antibody: AF488-conjugated AffiniPure alpaca anti-mouse IgG antibody (Jackson Immuno Research, 615-545-214)

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 mouse anti-human RelB antibody (antibodies-online, ABIN948396, lot 10307) diluted 1:50 in 1x Duolink Antibody diluent (Sigma-Aldrich, DUO82008) ON at 4°C.
• Wash cells 3x for 5min with PBS.
• Incubate cells with secondary AF488-conjugated AffiniPure alpaca anti-mouse IgG antibody (Jackson Immuno Research, 615-545-214) diluted 1:400 in 1x Duolink Antibody diluent for 1h 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 green staining dependent on the presence of ABIN948396 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)

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