PD-1 antibody (AA 201-288)

Catalog No. ABIN735608

There are 2+ publications for this product available. The Rabbit Polyclonal anti-PD-1 antibody is suitable to detect PD-1 in samples from Human, Mouse and Rat. It has been validated for FACS, WB, ELISA, IHC (p), IHC (fro), IF (cc) and IF (p). The antibody has been independently validated.

Quick Overview for PD-1 antibody (AA 201-288) (ABIN735608)

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

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 (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p))

Product Details anti-PD-1 Antibody

Binding Specificity: AA 201-288

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

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