CD84 antibody (Internal Region)

Catalog No. ABIN3187786

This anti-CD84 antibody is a Rabbit Polyclonal antibody detecting CD84 in WB, ELISA and IHC (p). Suitable for Human and Mouse. Independently validated for use in Immunohistochemistry.

Quick Overview for CD84 antibody (Internal Region) (ABIN3187786)

Target: CD84

Reactivity: Human, Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This CD84 antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Product Details anti-CD84 Antibody

Binding Specificity: Internal Region

Specificity: CD84 Polyclonal Antibody detects endogenous levels of CD84 protein.

Characteristics: Rabbit Polyclonal to CD84.

Purification: The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.

Immunogen: Synthesized peptide derived from the Internal region of human CD84.

Isotype: IgG

Application Details

Application Notes: WB 1:500-1:2000, IHC-P 1:100-300, ELISA 1:20000,

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104537 (Immunohistochemistry)

by: TRON

No.: #104537

Date: 12/07/2023

Antigen: CD84

Lot Number: 15601601

Method validated: Immunohistochemistry

Positive Control:

FFPE human tonsil

Notes:

Passed. The CD84 antibody ABIN3187786 specifically stains cells within the germinal zone in human tonsil.

Validation Images

IHC staining of human tonsil with anti-CD84 antibody ABIN3187786 (1:100) and counterstain with hematoxylin.

IHC staining of human tonsil with anti-PD-1 antibody ABIN3187786 (1:200) and counterstain with hematoxylin.

Full Methods

Primary Antibody: ABIN3187786

Secondary Antibody: HRP-conjugated goat anti-rabbit antibody (medac diagnostics, DPVR-110HRP, Lot 101122)

Full Protocol:

• Dewax and rehydrate tissue sections on glass slides in a descending xylene and graded alcohol series:
• • xylene 2x for 5 min.
  • 100 % (v/v) ethanol 2x for 5 min.
  • 96 % (v/v) ethanol 2x for 5 min.
  • 70 % (v/v) ethanol 2x for 5 min.
• Wash slides 2x 5 min with ddH2O.
• Cook slides for 30 min at 95 °C in citrate buffer pH 6.
• Wash slides 2x 3 min with ddH2O.
• Incubate slides in 0.3 % (v/v) H2O2 in PBS for 15 min to quench endogenous peroxidase.
• Wash slides 3x 3 min in 1X PBS.
• Block slides in 10 % goat serum in PBS for 30 min at RT.
• Incubate slides with primary rabbit anti-CD84 antibody (antibodies-online, ABIN3187786, lot 15601601) diluted 1:100 and 1:200 in 10 % goat serum in PBS ON at 4 °C in a humidified chamber.
• Wash slides 3x 3 min in 1X PBS.
• Incubate slides with secondary HRP-conjugated goat anti-rabbit antibody (medac diagnostics, DPVR-110HRP, Lot 101122) ready-to-use in 10 % goat serum in PBS for 30 min at RT in a humidified chamber.
• Wash slides 3x 3 min in 1X PBS.
• HRP development.
• Counterstain slides with hematoxylin solution (Carl Roth, T865.2).
• Wash slides for 15 min under running tap water.
• Dehydrate sections in an ascending alcohol-xylene series:
• • 70% ethanol 2x for 10 sec.
  • 96% ethanol 2x for 10 sec.
  • 100% ethanol 2x for 10 sec.
  • xylene 2x for 10 sec.
• Mount sections with Entellan new mounting medium (Sigma-Aldrich, 1008690500).
• Dry sections.
• Image acquisition on an Aperio CS2 Digital Pathology Slide Scanner (Leica), Magnification 40x.

Experimental Notes:

• Staining with ABIN3187786 overnight at 1:100 and 1:200 antibody dilution the signal in human tonsil was sharp and localized in the cells within the germinal zone (as expected). Modifying ABIN3187786 incubation conditions to 1 h at RT, the signal in human tonsil was still sharp and localized in the cells within the germinal zone.

• However, in mouse spleen, the signal was almost absent and very weak. With the modified incubation conditions, the signal in mouse spleen was again almost absent and very weak. With either staining protocol the signal in mouse lung was sporadic and its quality does not allow the possibility of non-specific staining to be excluded.

Handling

Format: Liquid

Concentration: 1 mg/mL

Buffer: Liquid in PBS containing 50 % glycerol, 0.5 % BSA and 0.02 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freeze/thaw cycles.

Storage: -20 °C

Storage Comment: Store at -20°C.

Target Details for CD84

Target: CD84

Alternative Name: CD84

Molecular Weight: 39 kDa

Gene ID: 8832

UniProt: Q9UIB8