PD-1 antibody (AA 201-288)

Catalog No. ABIN735608

Product Details anti-PD-1 Antibody

Target: PD-1 (PDCD1) (Programmed Cell Death 1 (PDCD1))

Binding Specificity: AA 201-288

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This PD-1 antibody is un-conjugated

Application: Flow Cytometry (FACS), Western Blotting (WB), ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunofluorescence (Cultured Cells) (IF (cc))

Cross-Reactivity: Human, Mouse, Rat

Predicted Reactivity: Cow,Horse,Rabbit

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human PD-1

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104535 (Immunohistochemistry)

by: TRON

No.: #104535

Date: 11/29/2023

Antigen: PD-1

Lot Number: BC04411027

Method validated: Immunohistochemistry

Positive Control:

FFPE human tonsil

Notes:

Passed. The PD-1 antibody (AA 201-288) ABIN735608 specifically stains cells within the germinal zone in human tonsil.

Validation Images

IHC staining of human tonsil with anti-PD-1 antibody ABIN735608 (1:250) and counterstain with hematoxylin.

IHC staining of human tonsil with anti-PD-1 antibody ABIN735608 (1:500) and counterstain with hematoxylin.

Full Methods

Primary Antibody: ABIN735608

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-PD-1 antibody (antibodies-online, ABIN735608, lot BC04411027) diluted 1:250 and 1:500 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 ABIN735608 the signal at 1:250 and 1:500 antibody dilution in human tonsil was sharp and localized in the cells within the germinal zone (as expected).

• ABIN735608 reactivity was also tested in mouse samples. However, in mouse spleen, the signal was almost absent and very weak at Ab dilution 1:250 and 1:500. No signal was detected in mouse pancreas. Even increasing the Ab concentration to 1:150 dilution, in mouse tumor tissue, ABIN735608 detected a very weak, unclear, and sporadic signal. The quality of the signal does not allow the possibility of background staining to be excluded.

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: ProClin

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

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-PD-1 antibody (ABIN735608)

Zhang, Niyazi, Hong, Tuluwengjiang, Zhang, Zhang, Su, Bao: "Effect of EBI3 on radiation-induced immunosuppression of cervical cancer HeLa cells by regulating Treg cells through PD-1/PD-L1 pathway." in: Tumour biology, Vol. 39, Issue 3, pp. 1010428317692240, (2017) (PubMed).

Szabó, Gáspár, Dajnoki, Papp, Fábos, Szegedi, Zeher: "Expansion of circulating follicular T helper cells associates with disease severity in childhood atopic dermatitis." in: Immunology letters, Vol. 189, pp. 101-108, (2017) (PubMed).

Target Details for PD-1

Target: PD-1 (PDCD1) (Programmed Cell Death 1 (PDCD1))

Alternative Name: PD-1/CD279 (PDCD1 Products)

Synonyms: CD279 antibody, PD-1 antibody, PD1 antibody, SLEB2 antibody, hPD-1 antibody, hPD-l antibody, Ly101 antibody, Pdc1 antibody, programmed cell death 1 antibody, PDCD1 antibody, Pdcd1 antibody

Background:

Synonyms: PD1, PD-1, CD279, SLEB2, hPD-1, hPD-l, hSLE1, Programmed cell death protein 1, Protein PD-1, PDCD1

Background: Inhibitory cell surface receptor involved in the regulation of T-cell function during immunity and tolerance. Upon ligand binding, inhibits T-cell effector functions in an antigen-specific manner. Possible cell death inducer, in association with other factors.

Gene ID: 5133

UniProt: Q15116

Pathways: Cancer Immune Checkpoints