CCDC42 antibody (Middle Region)

Catalog No. ABIN2785068

This anti-CCDC42 antibody is a Rabbit Polyclonal antibody detecting CCDC42 in WB. Suitable for Human, Mouse, Rat, Cow, Dog, Guinea Pig, Horse and Rabbit. Independently validated for use in Immunocytochemistry. This Primary Antibody has been cited in 2+ publications.

Quick Overview for CCDC42 antibody (Middle Region) (ABIN2785068)

Target: CCDC42 (Coiled-Coil Domain Containing 42 (CCDC42))

Reactivity: Human, Mouse, Rat, Cow, Dog, Guinea Pig, Horse, Rabbit

Host: Rabbit

Clonality: Polyclonal

Conjugate: This CCDC42 antibody is un-conjugated

Application: Western Blotting (WB)

Product Details anti-CCDC42 Antibody

Binding Specificity: Middle Region

Sequence: ENSEFEEIHE VIARYKTLVS MHHDLMQSAQ EGQEKIEHAK ARLARYMEEK

Predicted Reactivity: Cow: 100%, Dog: 100%, Guinea Pig: 100%, Horse: 100%, Human: 100%, Mouse: 100%, Rabbit: 93%, Rat: 93%

Characteristics: This is a rabbit polyclonal antibody against Ccdc42. It was validated on Western Blot.

Purification: Affinity Purified

Immunogen: The immunogen is a synthetic peptide directed towards the middle region of Rat Ccdc42

Application Details

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

Comment:

Antigen size: 316 AA

Restrictions: For Research Use only

Independent Validation (ICC)

Validation #103398 (Immunocytochemistry)

by: Developmental Biology, Johann-Friedrich-Blumenbach-Institute for Zoology and Anthropology, Georg-August-University of Göttingen

No.: #103398

Date: 10/12/2018

Antigen: CCDC42

Lot Number: QC28887-41199

Method validated: Immunocytochemistry

Positive Control: NIH/3T3 cells transfected with Cddc42-mCherry expression vector

Negative Control: Non-transfected NIH/3T3 cells

Notes:

Passed. ABIN2785068 specifically detects Ccdc42-Cherry when exogenously expressed. Background staining is low.

Validation Images

Cells were transfected with Ccdc42 tagged with mCherry (red) and incubated with ABIN2785068 (green). The pictures on the right show the red and green channels merged with DAPI counterstain (blue).

Full Methods

Primary Antibody: ABIN2785068

Secondary Antibody: goat anti-rabbit 488 antibody (MoBiotec, MFP-A1008, lot 2806F058)

Full Protocol:

• Grow NIH/3T3 cells (ATCC, CRL-1658) in DMEM+GlutaMAX (Gibco, 10567-014, lot 1922818) supplemented with fetal bovine serum (Gibco, 10500-064, lot 08FO477K) and Pen/Strep (Gibco, 15140-122, lot 1924797) at 37°C and 5% CO₂ to 70% confluency.
• Transfect cells with Ccdc42 tagged mCherry (Ccdc42-Cherry) expression vector using EndofectinMax (GeneCopoeia) following the manufacturer´s instructions.
• Grow cells for 24h.
• Fix cells in methanol for 10min at -20°C followed by incubation in 0.3% Triton X-100 for 10min at 4°C.
• Block cells in PBS containing 1% bovine serum albumin and 0.5% Tween-20 (PBT) for 1h at RT.
• Incubate cells with primary rabbit anti-Ccdc42 antibody (antibodies-online, ABIN2785068, QC28887-41199) diluted 1:100 in PBS for 1h at 37°C.
• Wash cells 3x 5min with 1x PBS.
• Incubate cells with secondary goat anti-rabbit 488 antibody (MoBiotec, MFP-A1008, lot 2806F058) diluted 1:300 for 1h at 37°C in PBS.
• DNA was counterstained with DAPI (4’, 6-Diamidino-2-phenylindole; Sigma, D-9542).
• Image acquisition on Zeiss LSM 510 confocal microscope and processing using Adobe Photoshop 5.0.

Handling

Format: Liquid

Concentration: Lot specific

Buffer: Liquid. Purified antibody supplied in 1x PBS buffer with 0.09 % (w/v) sodium azide and 2 % sucrose.

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: For short term use, store at 2-8°C up to 1 week. For long term storage, store at -20°C in small aliquots to prevent freeze-thaw cycles.

References for anti-CCDC42 antibody (ABIN2785068)

Wu, Yu, Liu, Wang, Huang, Lu, Chen, Li, Liu: "Essential Role of CFAP53 in Sperm Flagellum Biogenesis." in: Frontiers in cell and developmental biology, Vol. 9, pp. 676910, (2021) (PubMed).

Tapia Contreras, Hoyer-Fender: "CCDC42 Localizes to Manchette, HTCA and Tail and Interacts With ODF1 and ODF2 in the Formation of the Male Germ Cell Cytoskeleton." in: Frontiers in cell and developmental biology, Vol. 7, pp. 151 (PubMed).

Target Details for CCDC42

Target: CCDC42 (Coiled-Coil Domain Containing 42 (CCDC42))

Alternative Name: Ccdc42

Background: The function of this protein remains unknown.
Alias Symbols: RGD1565925
Protein Size: 316

Molecular Weight: 34 kDa

Gene ID: 303234

NCBI Accession: NM_001107009, NP_001100479

UniProt: D3ZAZ8